细胞生物学：endomembrane system 2014

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1、The structure of the cell is from the genes.is for the genes.is by the genes.is to the genes. (that government of the people, by the people, for the people) Endomembrane system and vesicular trafficThe major approaches of this chapter1.Terms on endomembrane system and organelles 2.The pathway of a s

2、ecretory protein formation3.The pathway of a membrane protein formation4.The pathway of lysosome formation5.Vesicles and vesicular traffic6.A special member of the endomembrane system (the peroxisome)7. Endomembrane system and diseasesThe major points of this chapter1.The biochemical components of e

3、ach organelles2.The structure and characteristics of each organelles3.The functions of each organelles4.The relationship and interaction between organelles Membrane organelles are related in generating and function inside the cell. Including nuclear membrane, endoplasmic reticulum, Golgi complex, ly

4、sosome and transport vesicles . Most people consider peroxisome as a special member of theendomembrane system. 1. Endomembrane system 内膜系统发生发展的关联：内膜系统发生发展的关联：The origination and development of endomebrane systemExtension of Outer membrane of nucleus Endoplasmic reticulum peroxisome Golgi complexPrim

5、ary LSSecondary LS Secretary vesicleMembrane protein vesicleA general prokaryotic cell A eukaryotic animal cellorganelle：structures in the cytoplasm with some chemical components, physical shape and special physiological functions . including :membrane organelles such as organelles in endomembrane s

6、ystem and mitochondra, non-membrane organelles such as ribosome, centrosome and cytoskeleton.inneroutercellCell membraneCytoplasm organelles Cytosol Membrane organellsNon-membrane organellsNucleus Endomembrane systemmitochondranucleolus ribosomecentrosomeproteasomecytoskeletonERGolgilysosomevesicleN

7、uclear matrixPeroxisome N. membranethe significance of the significance of endomembraneendomembrane system systemEnlarge the surface area for cell Enlarge the surface area for cell metabolismsmetabolisms内膜系统为细胞内生命活动提供了足够的表面积，内膜系统为细胞内生命活动提供了足够的表面积，大大提高了代谢效率。大大提高了代谢效率。Intracellular compartments for va

8、rious Intracellular compartments for various metabolism metabolism 内膜系统细胞器各自形成区域分隔，这种分隔保证内膜系统细胞器各自形成区域分隔，这种分隔保证了各细胞器独特的生化反应之间互不干扰了各细胞器独特的生化反应之间互不干扰。Specified function and coordinationSpecified function and coordination。构成内膜系统的各细胞器之间相互依存和协调，其构成内膜系统的各细胞器之间相互依存和协调，其中蛋白质的合成与分选是内膜系统的主要功能。中蛋白质的合成与分选是内膜系统的

9、主要功能。Attached ribosomesFree ribosomes2.The path of a secretory protein Secretive proteins ,membrane proteins and lysosome proteins share something in common. They are all synthesized in rough ER, and post-translational process are accomplished in Rough ER and Golgi Complex, the final products are so

10、rted in trans-Golgi net work and leave the GolgiComplex in the form of vesicle.The path of a secretory protein Cartoon 1 three kinds of proteins formation and transportation Endoplasmic reticulumMorphologymicrosomeMarker protein for ER : Glucose-6-phosphataseMicrosome vs microbodyClassfication of ER

11、According to the attached ribosomes：Rough ER（RER） （颗粒内质网（颗粒内质网,GER）Smooth ER（SER）（无颗粒内质网（无颗粒内质网,AER）The ratio of Rough ER vs Smooth ER varies from tissue to tissue, cell to cell. Cells like extrocrine cells (pancreatic cells) have more RER；muscle cells have more SER, hepatocytes have both。Biochemica

12、l components of ERlipidsGangliosides(神经节苷脂神经节苷脂)Plasmalogen(缩醛脂缩醛脂)Neutral lipid(中性脂中性脂)phospholipids 磷脂磷脂Phosphatidylcholine(卵磷脂卵磷脂55%)Phosphatidylethanolamine(磷脂酰乙醇胺磷脂酰乙醇胺20-25%)Phosphatidylinositol(磷脂酰肌醇磷脂酰肌醇5-10%)Phosphatidylserine(磷脂酰丝氨酸磷脂酰丝氨酸5-10%)Sphingomylin (鞘磷脂鞘磷脂4-7%) proeins 30-33条，条，Mr

13、15-150KDThe ratio of lipids and protein is 1：2proteinProteins involves detoxificationProteins involves lipid metabolismProteins involves carbohydrate metabolismcytP450, NADPH cytP-450 reductase, cytb5, cytb5 reductase,NADPH cytc reductase Fatty acid CoA ligase, Cholestrol hydroxylase , et al Glucose

14、-6- phosphatase（葡萄糖（葡萄糖-6-磷酸酶）磷酸酶）, -glucuronidase (-葡萄糖醛酸酶），glucuronyl transferase ，GDP-mannosyl transferase（甘露糖基转移酶）（甘露糖基转移酶）Retention proteinsChaperone protein classification of ERRough endoplasmic reticulum, RER Smooth endoplasmic reticulum ,SER Other specified ER structureattached with Ribosome

15、s Fine network of tubule structureMyeloid body(髓样体髓样体)Annulate lamellae(孔环状片层孔环状片层)Annulate lamellaeThe functions of rough ERRough ER is closely related to the synthesis of secretory protein, membrane protein and retention proteins for modification and transport.(1).it harbors ribosomes for vesicula

16、r transported proteinsProtein are synthesized in RER Secretory proteins(外输性的或分泌性的蛋白质外输性的或分泌性的蛋白质)Membrane integral protein（膜整合蛋白）（膜整合蛋白）Retention proteins （构成细胞器中驻留蛋（构成细胞器中驻留蛋白）（内质网，高尔基白）（内质网，高尔基体）体）Lysosomal proteins(溶酶体蛋白溶酶体蛋白)Signal peptide hypothesis How can a ribosome attach to RER, and how can

17、 a newly-made peptide be transported across the ER membrane into the chamber of ER? (2). Protein folding and modification in ERIn the lumen of ER：GSSG (oxidized glutathione GSSG ), protein disulfide isomerase, PDIheavy-chain binding protein, Bip ，which recognize misfolded protein and keep this prote

18、in in the lumen. Such as Heat Shock protein 70, HSP70.chaperone protein like calreticulin and glucose regulated protein 94, GRP94 the latter is endoplasmin, a marker Chaperone protein in ER these chaporone proteins share the same structure that they have the retentionSignal peptide - Lys-Asp-Glu-Leu

19、 (in short KDEL) Chaperone proteins that help newly synthesized peptide with proper disulfide linkage and folding. Chaprone proteinNewly-synthesized peptideNormal folded proteinMis-folded protein(3). Glycosylation (蛋白质的糖基化蛋白质的糖基化)Asn N-linked glycosylationTarget sequences: Asn-X-Ser or Asn-X-ThrAsn-

20、X- Ser or Asn-X-ThrThe roles of smooth ER(滑面内质网的功能滑面内质网的功能)（1）. Lipid synthesis（2）. Metabolism of glycogen（3）. Detoxification （4）. Regulation of Calcium concentration（5）. Gut acid and bile formation（1）. Synthesis of lipids Including: phospholipids ,cholesterol ,fat ,steroid hormones, glycolipids, et

21、 al. Smooth ER contains enzyme systems for lipids, phospholipids , cholesterol, and enzyme system for cholesterol transforming into its derivatives, such as various steroid hormones Cells secreting steroid hormones have more smooth EREndocrine cells ,such as genital gland and adrenal gland ,synthesi

22、ze steroid hormones in smooth ERLipids synthesis：most membrane lipids are synthesized in smooth ER（including phospholipids and cholesterol） two exceptions： sphingomylin and glycolipids（started at ER,finished,finished in in Golgi complex） Mit/Chl have some special lipids are made in their membrane. t

23、he ratio of phospholipids varies in different organelles： phosphatidylcholine（PC）：）： ERGCPM（highlow） phosphatidylserine（PS）：）： ER GC PM （lowhigh） three enzymes for phopholipids synthesis are located at ER as integral proteins. acylacyl transferasetransferase ( (脂酰基转移酶脂酰基转移酶) ) phosphotasephosphotase

24、( (磷脂酸酶磷脂酸酶) ) cholinecholine phosphotransferasephosphotransferase( (胆碱磷酸转移酶胆碱磷酸转移酶) )these enzymes have the activity on the cytoplasm side of ER(磷脂合成酶磷脂合成酶的的活性部位活性部位在在ERER膜的细胞质侧膜的细胞质侧) )that means the substrates of lipids synthesis are from cytoplasm (说明合成磷脂所需底物来源于细胞质基质说明合成磷脂所需底物来源于细胞质基质) )Basic re

25、action Basic reaction ：3 steps3 steps卵磷脂（磷脂酰胆碱）的合成过程：卵磷脂（磷脂酰胆碱）的合成过程： the fate of newly synthesized the fate of newly synthesized phospholipdsphospholipds（三种）（三种）flip-flop：新合成磷脂最初位于内质网膜的胞新合成磷脂最初位于内质网膜的胞质侧，部分磷脂在质侧，部分磷脂在磷脂转位因子磷脂转位因子（phospholipd translocator / flippase翻转酶）翻转酶）的作用下，将其从的作用下，将其从胞质侧翻转至胞质侧翻

26、转至内质网腔面内质网腔面。在在RER中的中的磷脂反转磷脂反转（A）ER membranecytosolER lumenNewly synthesized lipid added to the cytosol layerUnder the help of some Specific proteinDouble layers increased（B）cytoplasm membraneExtro-cellularintrocelluarFlopase help the plipidsWith flip-flopDouble layers increased transported to other

27、 organells as vesicles ：新合成磷新合成磷脂或者以出芽形成小脂或者以出芽形成小泡的形式运至泡的形式运至高尔基高尔基体、溶酶体、质膜体、溶酶体、质膜等。等。PhospholipidPhospholipid exchange exchange protinsprotinstransfer from ER to transfer from ER to other membrane by a other membrane by a carrier protein:carrier protein:With the help of phospholipid exchange prot

28、eins ，PEP, lipid will be translocated into other membrane。以。以。以。以形成复合物的形式自由扩散至细胞质基形成复合物的形式自由扩散至细胞质基形成复合物的形式自由扩散至细胞质基形成复合物的形式自由扩散至细胞质基质，然后运至质，然后运至质，然后运至质，然后运至线粒体、过氧化物酶体线粒体、过氧化物酶体线粒体、过氧化物酶体线粒体、过氧化物酶体等。等。等。等。phospholipid exchange proteins ：Hydrophilic proteins, are able to Hydrophilic proteins, are abl

29、e to associate with specific lipid and associate with specific lipid and transfer the lipid to other membrane transfer the lipid to other membrane （2）. The metabolims of glycogenlER involves in glycogen degradation： in hapetocyte , the SER in the cytosol layer (胞质面胞质面?) glycogen glycogen phosphoryla

30、seG-1-G-1-PP G-6-P，phosphotase in SER will remove the phosphate ，the glucose will be tranported into lumen of ER or transferred outside of cell membrane (circulation). lER involves in glycogen synthesis：it is controversial（3）. Detoxification the SER in hepatocytes has a lot of cyt b5, cyt P450，NADPH

31、-cytochrome reductase， NADH cyt b5 reductase et al electron transduction enzyme system. substrates will be oxidized or modified with OH, therefore, toxicity will be lost and will be moved out of our cells and body easily.（4）. Ca. Ca2+2+ storage and regulation storage and regulation As in muscle , it

32、 is specified as sarcoplasmic reticulum (肌细胞中的肌浆肌细胞中的肌浆网网特化的特化的SER)It invovles in regulation of calcium concentration in muscle contraction.( 参与肌原纤维收缩中的钙离子浓度调节参与肌原纤维收缩中的钙离子浓度调节)Releasing or storing calcium as Releasing or storing calcium as signaling molecules(signaling molecules(储存钙离子，作为细胞内信号物储存钙离子

33、，作为细胞内信号物质。质。) )（5）. Roles on synthesis and secretion of gut acid and bile (. Roles on synthesis and secretion of gut acid and bile (胃胃胃胃酸、胆汁的合成与分泌酸、胆汁的合成与分泌酸、胆汁的合成与分泌酸、胆汁的合成与分泌) ) 胃壁腺上皮细胞中，参与胃酸形成胃壁腺上皮细胞中，参与胃酸形成胃壁腺上皮细胞中，参与胃酸形成胃壁腺上皮细胞中，参与胃酸形成 肝细胞中参与胆汁的合成与分泌肝细胞中参与胆汁的合成与分泌肝细胞中参与胆汁的合成与分泌肝细胞中参与胆汁的合成与分泌Th

34、e Golgi Complexowl A City with lot of factoriesA harbor and post-processing centerMorphological types of Golgi ComplexMorphological types of Golgi Complex . .cisternacisterna( (扁平囊泡扁平囊泡) )： 3-8个个,弓形排列弓形排列(膜厚膜厚6-8nm，宽，宽15-20nm，囊间距，囊间距20 30nm ) .small vesicle .small vesicle（transport vesicletransport

35、vesicle运输囊泡运输囊泡）：）：（ 404080nm 80nm ） .large vesicle.large vesicle（secretorysecretory vesicelevesicele分泌泡）：分泌泡）：（ 10010050500nm0nm）3D structure of Golgi complex3D structure of Golgi complexMedial region: Medial region: medial medial cisternacisternaTrans region including: Trans region including: secr

36、etorysecretory vesicles, trans vesicles, trans cisternacisterna and and trans Golgi network,TGNcis region including : transferred vesicles , cis Golgi network(CGN ) and cis cisterna Distribution Distribution lShape , distribution and physiological status are closely related with cell types and cell

37、functions.(形态、分布、生理状态与细胞类型形态、分布、生理状态与细胞类型及功能有关；及功能有关；)lThe abundance of Golgi Complex is closely related to the differentiation of the cell (发达程度与细胞分化程度呈正比；发达程度与细胞分化程度呈正比；)lThe location of Golgi Complex with The location of Golgi Complex with cytoskeleton(cytoskeleton(高尔基体与细胞高尔基体与细胞骨架关系密切骨架关系密切) )：

38、in non-polar cell, Golgi Complex locates at negative part of in non-polar cell, Golgi Complex locates at negative part of MTOCMTOC ( (在非极性细胞中，高尔基体分布在在非极性细胞中，高尔基体分布在MTOC(MTOC(负端负端) ) ) there are motor proteins on there are motor proteins on cistenacistena membrane of Golgi Complex. membrane of Golgi

39、Complex. 高尔基高尔基体体的膜囊上存在微管的的膜囊上存在微管的马达蛋白马达蛋白（cytoplasmic dynein和和kinesin）和微丝的和微丝的马达蛋白马达蛋白（myosin）。 Chemical components of Golgi Complex(高尔基复合体的化学组成高尔基复合体的化学组成)llipid：in between of cytoplamic membrane and ER，40%； lprotein：60% enzyems：many kinds，such as：glycosyltransferase 、phospholipase , oxidoreducta

40、se(磷脂 酶、氧化还原酶)(most in trans-)；lSugar chain ：increasing from cis to trans(多糖分布从形成面到成熟面呈梯度 上升；)Lipid type ER Golgi Complex cell membraneTotal 30 35 42Phospholipid 26.6 28.5 29.8SM 1.3 3.5 5.6PC 13.5 14.6 14.0PE 6.4 6.6 6.9PI 2.1 2.6 2.1CHO 1.0 1.7 5.2glycolipid 0.4 1.5 2.5Comparison of the lipid in E

41、R, Gogi complex, cytoplasm membrane in rat hepatocytes（%） type functionstype functionsglycosyltransferase involves glycoprotein formation （Marker） manose transferase N-acetylgalactose transferase N-acetylglucosamine transferase N-acytylneuraminic acid transferase galactose transferase Enzymes in Gol

42、gi ComplexCis-cisternaTrans cisternaMedial cisternaThe biological roles of Golgi Complex（1）it involves in protein modification ,transportation , sorting and secretion. (外分泌性蛋白、细胞膜外分泌性蛋白、细胞膜 蛋白、溶酶体酶蛋白、蛋白、溶酶体酶蛋白、细胞外基质蛋白等都通细胞外基质蛋白等都通高尔基体修饰和转运高尔基体修饰和转运)Golgi Complex provide the sites for protein and lip

43、id modification glycoprotein glycoprotein glycosylationglycosylation ( (糖蛋白的糖基化糖蛋白的糖基化) ) glycolipidglycolipid glycosylationglycosylation ( (糖脂的糖基化糖脂的糖基化) )Protein Protein glycosylationglycosylation( ( 糖基化修饰的蛋白有两种连接方式糖基化修饰的蛋白有两种连接方式) )： N Nlinkedlinked： N N连接在连接在ERER腔内完成腔内完成，仍需在高尔基复合体内进一步加工修饰（剪切仍需在高

44、尔基复合体内进一步加工修饰（剪切和补加某些基团）和补加某些基团） Olinked： O连接主要发生在连接主要发生在Golgi complex与与Ser、Thr和和Hyp的的OH连接，连接的糖为半乳糖或连接，连接的糖为半乳糖或N-乙酰半乳糖胺。乙酰半乳糖胺。 protein glycosylation(N-linkage vs O-linkage)characteristics N-linkage o-linkage1. Formation site RERGolgi Complex2. Ways of formation from an oligo-glucose groupAdded one

45、 after another3. Amino acid AsnSer、Thr、4 Length at least five sugarMost 1-6 sugar，Some may be longer, ABO5.The first sugarNacetylglucosaminegalactose, NacetylgalactosamineThe core carbohydrate of N-linked oligosaccharides is assembled in the rER. Modifications to N-linked oligosaccharides are comple

46、ted in the Golgi complex. O-linked oligosaccharides takes place in Golgi complex.The characteristics of glycosylation in Golgi ComplextThe enzymes for glycosylation in Golgi complex are integral proteins(高尔基体的糖基化修饰加工酶都是整合膜蛋白；)tEnzyme activity differs from one part to another- compartmentalization (定

47、位区域不同酶加工区室化；)tEnzyme activity in inner side of cisterna membrane(加工酶的活性部位面向高尔基体腔面；)tEnzyme activity including glycosidase and glycosyl transferase(作用：切去或加上单糖基；)The role of The role of glycosylationglycosylationprotecting protein from damage or digestion of proteinase (对蛋白质有保护作用,使蛋白质能够抵抗消化酶的作用)；provi

48、ding recognition signal for cell-cell recognition(赋予蛋白质运输信号的功能)；Even providing the signal for some protein to fold(某些蛋白只有在糖基化之后才能正确折叠。)involving cell surface coat or glycocalyx (参与形成细胞表面的糖被.)(1(1)In )In GolgiGolgi Complex, Complex, inactive inactive precursor precursor protein protein will will be b

49、e cleaved into active form , such as insulincleaved into active form , such as insulin (2)Some (2)Some neurotransmitter neurotransmitter (peptides) (peptides) share share the the same same precursor, precursor, in in GolgiGolgi complex, complex, this this precursor precursor will will be be cleaved

50、cleaved into into several several peptides.peptides.(3)In (3)In GolgiGolgi complex, complex, sulfate sulfate group group will will be be added added in in TyrTyr site site at OH, later on will form at OH, later on will form proteoglycanproteoglycan( (蛋白聚糖蛋白聚糖) )； （2 2）.involving post-protein synthes

51、is .involving post-protein synthesis modification or digestion and maturation (modification or digestion and maturation (参与蛋参与蛋白质的水解加工白质的水解加工) )Post modification of insulinERGolgi Coplexcytosollumenadreno-cortico-tropic-hormone 蛋白质由RER合成、修饰后在高尔基体内分选，后再由运输小泡送往细胞的不同靶结构部位。Base for sorting(分选基础)：glycopr

52、otein has special modification or mark (糖蛋白的修饰具有方向性，使蛋白具有特异的分选标志)；mannose-6-phosphate(M-6-p) is a special mark for lysosome sorting(甘露糖基磷酸化也是分选的标签)。The role of Golgi complex here(高尔基体作用)：the transition and docking part (膜泡运输的重要枢纽)；（3）involving protein sorting and vesicle transport(参与蛋白质的分选与胞内参与蛋白质的分

53、选与胞内囊泡运输囊泡运输)形式：形式：形成运输小泡或分泌小泡形成运输小泡或分泌小泡3.The process of membrane protein formation (膜蛋白的形成过程膜蛋白的形成过程)Nucear proteinsParts of Mit proteinsPeroxisome proteinsProteins in cytosolSecretion proteinsMembrane integral proeinsRetention proteins Lysosomal enzyme proteinsThe process of lysosome formation(溶酶

54、体的形成过程溶酶体的形成过程)The lysosomal protein synthesis, modification,Sorting and transporting(溶酶体内蛋白的形成溶酶体内蛋白的形成)The characteristics of lyososome structure and functions溶酶体结构功能特点溶酶体结构功能特点lysosomeOne One unit-membrane unit-membrane enclosed enclosed structure, structure, contains contains many many kinds kin

55、ds of of acid acid hydrolaseshydrolases for for cellular cellular digestion digestion and and recycling recycling ( ( 它它是是单单层层膜膜围围绕绕、内内含含多多种种酸酸性性水水解解酶酶类类的的囊囊泡泡状状细细胞胞器器，其其主主要要功功能能是是进进行细胞内消化行细胞内消化) )。heterogenous，in size and different kinds of hydrolasesMorphological character and Morphological charac

56、ter and chemical components(chemical components(溶酶体溶酶体的的形态形态结构和化学组成结构和化学组成) )From 0.2-0.8mevenfrom 0.05m to couple mMorphology characteristics.mono-membrane, sphere like. contains： enzymes enzymes for for digestion, digestion, belong belong to to acid acid hydrolaseshydrolases, , optimal pH is 5.0.

57、optimal pH is 5.0. Marker Marker enzymeenzyme：acid phosphatase小鼠腹腔巨噬细胞小鼠腹腔巨噬细胞The membrane proteins of lysosomesATPADP+PiH+The process of lysosomal protein synthesis , modification,sorting and transportingLysosomalproteinsMembrane proteins（highly glycosylated to protect fromEnzyme digestion, and to

58、protect the membrane.）The formation and process are similar to cytoplasm Membrane proteins.Lysosomal enzyme proteins ,activation at pH 5.0 the formation and process are similar to secretion Proteins, but in Golgi , a marker M-6-P will added for sorting.Integral membrane proteins of lysosomesA. Integ

59、ral proteins highly glycosylated for protection from digestion(溶酶体膜蛋白高度糖溶酶体膜蛋白高度糖基化基化，可有利于防止自身膜蛋白的降解，可有利于防止自身膜蛋白的降解)。如如lgpA、lgpBB. Membrane with a lot of H+ pump to main a acidic environment(上上嵌有质子泵嵌有质子泵，形成和维持，形成和维持溶酶体中酸性的内环境溶酶体中酸性的内环境)； C. membrane with many kinds of carrier protein for transport t

60、he digested products for recycle or reuse.（并且并且具有具有多种载体蛋白多种载体蛋白，用于水解的产物向外转运用于水解的产物向外转运；） the homology of lysosomal membrane proteins(溶酶体膜糖溶酶体膜糖蛋白家族的同源性蛋白家族的同源性)lysosomal-associated membrane protein，LAMP lysosomal integral membrane protein，LIMPInclude: N-signal peptide lumen domain（highly glycosylate

61、d） Transmembrane（uni-transmembrane） C-cytosol domain（10AAs，highly conservative, might be related as sorting signal ） The process of digestive enzyme formation in lososome(溶酶体水解溶酶体水解酶的形成酶的形成)（1 1）. Enzyme proteins are synthesized and . Enzyme proteins are synthesized and folded , N-folded , N-glycosy

62、latedglycosylated and and translocatedtranslocated from ERfrom ER（2）. Lysosomal enzyme protein are modified and transported in Golgi Complex modification : glycosylation , phosphorylation .in cis face or network：marked with mannose-6-phosphate,M6P N-acetyglucosamine phosphotransferase （3）. Lysosomal

63、 protein sorting and transporting in tans face of Golgi, there are receptors for M-6-P，budding with clathrin coated visicle，leave Golgi。（4）. Primary lysosome formation Uncoated visicle forms, under acidic conditions, lysosomal protein dephosphate, M-6-P receptors recycle to transface of Golgi Comple

64、x寡糖链寡糖链溶酶体酶蛋白溶酶体酶蛋白甘露糖甘露糖-6-6-磷酸受体磷酸受体The recognition of a lysosomal hydrolase. The GlcNAc phosphotransferase enzyme that recognizes lysosomal hydrolases in the Golgi apparatus has separate catalytic and recognition sites. The catalytic site binds both high-mannose N-linked oligosaccharides and UDP-

65、GlcNAc. The recognition site binds to a signal patch that is present only on the surface of lysosomal hydrolases. 溶酶体分选的信号斑溶酶体分选的信号斑Types of lososomesClassified according to their function primary lysosomesecondary lysosometertiary lysosome post- lysosome residual body telolysosome（1 1）. . （primary

66、lysosome）lNewly secreted from Newly secreted from GolgiGolgi Complex, no Complex, no enzyme activity enzyme activity小鼠脾脏巨噬细胞中的溶酶体小鼠脾脏巨噬细胞中的溶酶体用电镜细胞化学技术显示其中含有的酸性磷酸酶。用电镜细胞化学技术显示其中含有的酸性磷酸酶。 M：线粒体，线粒体，L：溶酶体（朴英杰）溶酶体（朴英杰）（2 2）. secondary . secondary lososomelososomel primary primary lososomelososome merge

67、 with merge with phagosomesphagosomes or or autophgosomeautophgosome and trigger the proton pump , pH and trigger the proton pump , pH will drop to 5.0, the enzymes will be active. will drop to 5.0, the enzymes will be active. Phagolysosome autophgolysosomeAccording to the origin of substratesAccord

68、ing to the origin of substrates autophagic lysosome heterophagic lysosome Phagocytosis and Autophagy（3 3）tertiary lysosomepost-lysosome，no enzyme activity Also called Also called residulresidul body or body or telolysosomeThe functions of lysosomes（1） digestive digestive oranelleoranelle: to recycle

69、 the old, used , : to recycle the old, used , organelles as energy or nutrition. organelles as energy or nutrition. to digest some components for energy, such as LDL- to digest some components for energy, such as LDL-ChOChO（2 2） protective protective functionsfunctions（病原体感染刺激单核细胞（病原体感染刺激单核细胞分化成巨噬细胞

70、而吞噬、消化分化成巨噬细胞而吞噬、消化） 过程过程： phagocytosis phagosomeendocytosis early endosome late endosomelysosome autophagy autophagosome Digestion of bacteria or virus （3） involving some homone metabolism （4） involving development (在个体发生、发育在个体发生、发育过程起重要作用过程起重要作用) involve in clearance of extro or degenitive tissues

71、 of cells . Bone develop or remodeling Mammal uterus cycle , human menstruation fertilization （acrosome reaction)Cellular vesicles and vesicle transportCellular transportation system细胞内的运载系统（囊泡运输细胞内的运载系统（囊泡运输）Types of coated vesicles and oriented transport (有被囊泡的种类及定向转运有被囊泡的种类及定向转运)127vesicle128Vesi

72、cle transport129types and characteristics of vesicle transport(囊泡运输的类型及囊泡运输的类型及特点特点) normal transporting ways among endosystem parts such as :ERGolgi；Golgilysosome； secretion particle extrocelluar part.target organelles have specific membrane marker.vesicle transporting move along microtubles or mic

73、rofilements , motion power from motor proteins.130Generating and types of vesicles(囊泡的来源与类型囊泡的来源与类型) Most vesicles formation via budding at a specific area on a membrane structure. The surface area of the vesicles has coat, which will be disassembled before merging with target membrane. The roles of

74、 the coat of vesicles(衣被的主要作用衣被的主要作用衣被的主要作用衣被的主要作用) )：selectively packaging specific proteins to form vesicles；driven force for budding determine the orientation of the vesicles131 3 kinds coated vesicels（1）. Clathrin coated （2）. COPII coated（3）. COPI coated133clathrin-coated vesicle134Selective tra

75、nsport by clathrin coated vesicles135componentscomponents： 1 1）adaptor proteinsadaptor proteins： AP1：Golgiendosome AP2：cytoplasm membraneendosome AP3：Golgilysosome 1362 2）dynamin（发动蛋白，缢断蛋白）发动蛋白，缢断蛋白）Hydrophilic proteinHydrophilic protein，a kind of a kind of GTPaseGTPase。137Clathrin coated vesicle fo

76、rmation138COP coated vesiclestransport from ER to Golgi （cis-tranportation）；）； COP contains 5 subunits，sec12p、 sec24p、sec23p、sec13p、sec31p； assembly controlled by Sar protein；COP coated vesicle recognize specific protein and can concentrate the proteinFor ： COPCOPCOPCOP coating protein recognizes th

77、e transmembrane receptor of the cargo receptor, while the receptor binds to the specific cargo. COP coated vesicle is a temporary structure ,will de-coat before its merging with the target.140COP包被小泡包被小泡 (COPcoated vesicles)responsible for resending proteins escaped from ER to Golgi, (escaped protei

78、ns） ER。 COPcontains 8 subunits， assembly or dis-assembly depends on ARF a GTP-binding protein. 141COPI衣被小泡衣被小泡142the mechanism for organelles to keep or recycle or retention proteins： a. a. before before the the coated coated vesicle vesicle formation, formation, most most retention protein are excl

79、uded from budding.retention protein are excluded from budding. b.b. through through specific specific recognition recognition signal signal peptide peptide , ,lys-lys-asp-glu-leu,KDELasp-glu-leu,KDEL ， and and receptor receptor association, association, COPCOPcoatedcoated vesicle vesicle will will c

80、o-send co-send back back the the escaped escaped protein from protein from GolgiGolgi to ER. to ER.。 143内质网驻留蛋白的回收图解内质网驻留蛋白的回收图解Types of coated vesicles and their functionsER to ER to GolgiSec23/Sec24 complexcomplex，Sec 13/31complexcomplex， Sec 12Sar 1COP IIGolgiGolgi to ER to ERCOPARFCOP IPlasmPlas

81、m membrane to membrane to endosomeendosomeGolgiGolgi to to endosomeendosomeGolgiGolgilysosomelysosomeClathrin H and L chain H and L chain，AP2Clathrin H and L chain H and L chain，AP1Clathrin H and L chain H and L chain，AP3ARFclathrinTransportation Transportation dircectiondircectionComponents and Com

82、ponents and adpatoradpatorproteinsproteins GTP binding binding proteinproteintypetype146vesicular transport（1）vesicle transport is the basic way for material transport with proper orientationVesicular transport：vesicle budding from a organelle and leave that organelle ,then transported to a target o

83、rganelle and merged with the target organelle.147（2）vesicular tranport is a highly ordered , strictly selected and precisely controlled process for cargo transportation.Selcetion of pathway or route Selection of the cargo, even quality, folding modification.148（3 3）specific recognition guarantees th

84、e specific recognition guarantees the precise dockingprecise dockinglSpecific recognition and merging。lCoated vesicles move along some path to target, motor Coated vesicles move along some path to target, motor protein and ATP will provide energyprotein and ATP will provide energylAll vesicles are a

85、ble to merge with target precisely, All vesicles are able to merge with target precisely, because of vesicle has its marker protein which can be because of vesicle has its marker protein which can be recognized by target receptors. Two proteins are recognized by target receptors. Two proteins are im

86、portant for cargo vesicle be recognized by target. important for cargo vesicle be recognized by target. SNAREs（soluble NSF attachment protein receptor）：）：Rabs（targeting GTPase）：其作用是使运输小泡靠近靶膜，其作用是使运输小泡靠近靶膜，催化融合的发生。催化融合的发生。anchor 150.SNAREs Functions ：mediate cargo vesicle merge with target。types：20 k

87、inds of SNAREs were found in animals， distribute in membrane。v-SNAREs：运输小泡膜上运输小泡膜上（囊泡相关膜蛋白）（囊泡相关膜蛋白）t-SNAREs：靶膜上：靶膜上（联接蛋白）（联接蛋白）转运小泡与靶向小泡或膜式结构通过转运小泡与靶向小泡或膜式结构通过v-SNARE 与与t-SNARE 识别融合的示意图识别融合的示意图（1）运输囊泡停泊）运输囊泡停泊v-SNAREt-SNARE(2) t-SNARE与与v-SNARE识别识别（3）钙离子从钙离子通道释放）钙离子从钙离子通道释放（4）钙离子与）钙离子与SNARE的结合促进囊泡与靶

88、膜的融合的结合促进囊泡与靶膜的融合154structure：has helix structureActivation ：SNARE molecules need to be released from binding state (reuse, or recycle) NSF is a molecule like chaprone ,a ATPase，by using the energy from ATP, makes adaptor protein insert into SNAREs complex to separate snares.155SNAREs in vesicle tra

89、nsport（ T-SNARE ， V-SNARE ）156SNARE was separated by NSF and adaptor proteins158（4 4）vesicular transport is the bridge for vesicular transport is the bridge for membrane structure transition and metabolism membrane structure transition and metabolism interchanginginterchangingmembrane flow：the membr

90、ane portion may translocate , merge, exchange or recombinate among endomembrane system or organelles or cytoplasm membrane. this membrane flow is not only for cargo transport, but also for membrane renew.Golgi Complex is the transportation junction of the vesicular Transport. 159Types of Vesicle Tra

91、nsport and Their FunctionsA. Three different types of coated vesicles.Different coat proteins select different cargo and shape the transport vesicles that mediate the various steps in the biosynthetic-secretory and endocytic pathways.Dynamic Membrane Flow Through the CellSummary of Endomembrane Syst

92、emvEndomembrane System : The structural and functional relationship organelles including ER,Golgi complex, lysosome, endosomes, secretory vesicles.vMembrane-bound structures (organelles) are found in all eukaryotic cells.A special organelle in endomembrane system,the peroxisomeAnother detoxification

93、 organelle a a one one layer layer of of unit unit membrane membrane enclosed enclosed body body with with enzymes. enzymes. heterogenous 。 1954，J. Rhodin fist discovered in mouse kidney epithelia of renal tubule. .Peroxisome or microbodyBiochemical components40 enzymes have been found40 enzymes hav

94、e been found1 1）. .oxidaseoxidase ( (氧化酶类氧化酶类): such urate oxidase 、L-amino L-amino acid acid oxidaseoxidase、D-amino D-amino acid acid oxidaseoxidase et et al. al. flavinflavin dependent dependent oxidaseoxidase2 2）. .catalasecatalase ( (过氧化氢酶过氧化氢酶) ) Marker enzyme for Marker enzyme for peroxisomepe

95、roxisome3 3）. . Peroxidase (过氧化物酶过氧化物酶) )Function of peroxisome 1）. Detoxification largely exists in animal hepatocytes and kidney cells two major enzymes for detoxificationtwo major enzymes for detoxification FAD dependent FAD dependent oxidaseoxidase to produce to produce H H2 2O O2 2 RH2+O2 R +H2

96、O2 catalasecatalase：to make to make H H2 2O O2 2 form waterform water RH2+H2O2R+2H2O 2 2）. . Regulation Regulation of of oxygen oxygen concentration(concentration(对对氧氧浓度的调节作用浓度的调节作用) )： to protect cell from high oxygen toxicityto protect cell from high oxygen toxicity3. Oxidation of fatty acid(脂肪酸脂肪

97、酸的的氧化氧化) )： 过氧化物酶体过氧化物酶体中有中有氧化脂肪酸的酶类，可氧化脂肪酸的酶类，可分解分解 脂肪酸等高能分子向细胞直接提供热能脂肪酸等高能分子向细胞直接提供热能。4. Metabolism of molecules with nitrogen(含含氮氮物物质的代谢质的代谢)： 尿酸氧化酶可将核苷酸和某些蛋白质降解代谢尿酸氧化酶可将核苷酸和某些蛋白质降解代谢 的产物进一步氧化去除。的产物进一步氧化去除。Generation of Generation of peroxisome(peroxisome(过氧过氧化物酶体的化物酶体的发生发生) ) Peroxisomal-targeti

98、ng signal，PTS）：）： PTS1 :Ser-lys-leu，in C terminalin C terminal PTS2: Arg/Lys-Leu/lle-5X-His/Gln-leu，in N-terminalin N-terminal membrane protein as receptors to membrane protein as receptors to recoginizerecoginize PTS PTS 1.Direct division to form new peroxisomes, enzymes will be moved in for matura

99、tion 2 Membrane directly from ER, enzymes are moved into for maturation Enzymes of perosisome are encoded by nuclear DNA,synthesized in cytosol, transported into peroxisome 过氧化物酶体发生过程的示意图过氧化物酶体发生过程的示意图the differences between lysosome and peroxisome 1.Similar size and morhphology. But peroxisome has

100、high concentration of urate oxidase, even form crystal, a special property for EM 2. Separate them by centrifigution 3. Enzyme difference4. Origin in different waysComparison of lysosome and peroxisome Road map of protein sortingRoad map of protein sorting Endomembrane system and diseases内膜系统的任何一部分的

101、结构和功能的异常都导致疾病的发生。内膜系统的任何一部分的结构和功能的异常都导致疾病的发生。181ER and diseases ER a sensitive organelleER a sensitive organelle pathological changespathological changes：Swollen under adverse conditions such as hypoxia ,irradiation, toxicity or infection, water and sodium will move into ER lumen. ER lumen. Loss of

102、ribosomeAccumulation of materials such as unfolded protein.Tumor cell , ER might have morphological changes such as annulate lamellaeAnnulate lamellae183 eg：1-antitrypsin(1-AT) (肝细胞中合成的糖蛋白肝细胞中合成的糖蛋白) Deficiency：blood lack 1-AT, while 1-AT accumulated in RERcause：mutation of 1-AT happened ,so that 1-

103、AT is not able to transport to bloodClinic relevance ：new born baby hepatitis ，emphysema 1-AT is a inflammation protein, is able to protect cells from proteinase attack. 184Golgi Complex and diseases1）、）、enlargement ：hyper-function2）、）、withered or damaged: toxicity 3）、）、content changes4）、）、correlati

104、on with differentiation： positiveTumor cells have low Golgi complex ratioLysosomeLysosome and diseases and diseases Causes：abnormal enzyme、stability of membrane decreased1 1）. Genomic diseases. Genomic diseases：lack enzyme or enzyme activitylack enzyme or enzyme activity exhibition ： material over-l

105、oaded such as lipid deposit disease, glucogen deposit diease.Type 2 glycogenosis, pompers diseaselack a-glucosidaseheart,liver,kidney,muscle.Gaucher abnormal glucosidase；NiemannPick）lacksphingomyelinase186台台-萨氏（萨氏（Tay-Sachs）（黑朦性先天愚病）神经元中的溶酶体）（黑朦性先天愚病）神经元中的溶酶体lTay-Sachs（黑朦性先天愚病）（黑朦性先天愚病） N- acetyl gl

106、ucosidase AGM2 accumulationloss sight 、num、paralysis ；187Silicosis ：damage of lysosomal membraneOccupational disease；inhalation of crystalline silica dust （SiO2）, macrophage catch the dust by endocytosis, negatively charged SiO2 associate with lysosomal membrane, the lysosomal membrane is damaged, l

107、eads cell to necrosis , further damage other cells, results in fiber formation, function loss treatment：poly-2-vinylpyridine-N-oxide (PVNO,P-204) bind to SiO2 ,to protect lysosomal membrane2）. Diseases from damage: non-genetic diseases188Gout Gout is a disorder of purine metabolism,8 and occurs when

108、 its final metabolite, uric acid, crystallizes in the form of monosodium urate, precipitating in joints, on tendons, and in the surrounding tissues.6 These crystals then trigger a local immune-mediated inflammatory reaction. Uric acid is able to form hydrogen bond with lysosomal membrane ,which resu

109、lts in instability of lysosome, crystal of uric acid will damage lysosome , therefore, break the cell and incurs inflammation.1891）. Genetic diseases caused by lack enzyme or enzyme activitylCatalase loss： no catalase blood lEmpty peroxisome ：genetic mutation , lack receptors for enzyme transport。Ze

110、llweger syndrome ：遗传性疾病，与酶蛋白输入有关的蛋：遗传性疾病，与酶蛋白输入有关的蛋白质变异，过氧化物酶体空载白质变异，过氧化物酶体空载.2）. Pathological changes of peroxisomelAlcohol toxicity, virus mediated hepatitis, peroxisome number increasedltumor：peroxisome number decreased；Peroxisome and diseasesTerms or names：endomembrane system,microsome,signal pe

111、ptide,signal peptide recognition particle，translocator, signal peptide theoryquestions：1.How many kinds of protein are synthesized ,transported and modified in RER? please explain each step of these process.2. Please write down how many kinds of glycosylation, tell where they take place and what are

112、 the function of glycosylation. 3.Eplain the function of ER. What is the marker enzyme of ER? Endomembrane system and ERquestions1、what is the characteristics of Golgi Complex in morphology,what is the function of Golgi Complex? 2、Briefly explain types of protein glycosylation, and their characteris

113、tics and sites of formation. 3、How can Golgi Complex sort protein? Give an example.4、what is the marker enzyme of Golgi Complex?Golgi ComplexTerms：primary lysosome, secondary lysosome, tertiary lysosome(post- lysosome, residual body telolysosome), Phagolysosome, autophagic lysosome ， heterophagic ly

114、sosome, Questions 1、how many kinds of lysosomes?2、how dose lysosome form ? What are functions of lysosomes?3、How do lysosomes digest materials from outside the cell and inside the cells? 4、what are the marker enzymes of lysosome and peroxisome? What are the major differences between the two organell

115、es? 5、why do call lysosome and peroxisome heterogenous organelles?6、what are the functions of peroxisome? Compare the detoxification function of peroxisome and smooth ER.lysosomes193Terms Terms vesicle transportvesicle transport，COPII coated vesicleCOPII coated vesicle，COPI COPI coated vesicle, gene

116、tic coated vesicle, genetic lysosomelysosome diseases diseases Questions Questions 1.how many kinds of major vesicles? what are the 1.how many kinds of major vesicles? what are the transports mediated by them, and what are the transports mediated by them, and what are the orientations?orientations?

117、2.what are the significance of vesicle transport? 2.what are the significance of vesicle transport? 3.explain the mechanism of vesicle transport has 3.explain the mechanism of vesicle transport has the proper direction.the proper direction. 4.what is the pathological mechanism of silicosis ?Vesicle transport