1、Chapter 6 Cytoplasmic matrix, Endomembrane system, Protein Sorting and membrane traffickingLearning objective1. Compartmentalization in Eukaryotic Cells; The structural and functional relationship among the ER, Golgi complexes, lysosomes and plasma membranes of eukaryotic cells;The pathways of prote

2、ins targeting and sorting, and its mechanisms; The ways of protein modifications and intracellular sites after they are synthesized;Types of vesicle transport and their functions.誣麓銷沫廚瞞瞄耳埔跡臥挫堪掃回覽廚侈棄刨魏簇造波屏霖慧頁呸雄皇嚴考研資料：北京大學細胞生物學 第六章 基質與內膜(上)考研資料：北京大學細胞生物學 第六章 基質與內膜(上)Chapter 6 Cytoplasmic matrix1. The

3、Compartmentalization in Eukaryotic CellsMembranes divide the cytoplasm of eukaryotic cells into distinct compartments. Three categories in eukaryotic cells: (1) the endomembrane system: ER, Golgi complex, Lys., secretory vesicles. (2) the cytosol. (3) mitochondria, chloroplasts, peroxisomes, and the

4、 nucleus. Membrane-bound structures (organelles) are found in all eukaryotic cells.默顏訟蕾揍合姥幼持涅殿碧丑歹調豢累搽盅咖震戎韭息味佩增踞撐妙啃婁考研資料：北京大學細胞生物學 第六章 基質與內膜(上)考研資料：北京大學細胞生物學 第六章 基質與內膜(上)1. The Compartmentalization inCytoplasmic matrix and its functionsCytoplasmic Matrix: The region of fluid content of the cytoplasm

5、outside of the membranous organelles. Aqueous solution of large and small molecules including filaments of cytoskeleton which act as organizer for some order.The Cytosol is the site of protein synthesis and degradation or modification. It also performs most of the cells intermediary metabolism.Cytop

6、lasmic matrix (Cytosol) and Endomembrane System礫躊聯茸匪綴竿翠唱吊恐佬號套閉聳空登稽妻腮磐鴛綸婚毯局蜀胖鈕郡棲考研資料：北京大學細胞生物學 第六章 基質與內膜(上)考研資料：北京大學細胞生物學 第六章 基質與內膜(上)Cytoplasmic matrix and its funFunctions of cytoplasmic matrix:The protein synthesis, degradation and modification.Cells carefully monitor the amount of misfolded prote

7、ins. An accumulation of misfolded proteins in the cytosol triggers a heat-shock response, which stimulates the transcription of genes encoding cytosolic chaperones that help to refold the proteins.臥贓堅連磐惶兵遣冤脊訝稠約愉烈鶴曠堂鵝回劫汪撼洶致稻徒狹陋危袖刷考研資料：北京大學細胞生物學 第六章 基質與內膜(上)考研資料：北京大學細胞生物學 第六章 基質與內膜(上)Functions of cyto

8、plasmic matri搞箔付鮑袖輾瞬陪霉桐窺穴疵應憊烏藻收脆跨饑蘑駒駛匹旁漬群隱榜近蒼考研資料：北京大學細胞生物學 第六章 基質與內膜(上)考研資料：北京大學細胞生物學 第六章 基質與內膜(上)搞箔付鮑袖輾瞬陪霉桐窺穴疵應憊烏藻收脆跨饑蘑駒駛匹旁漬群隱榜B. Endomembrane SystemEndomembrane System : The structural and functional relationship organelles including ER,Golgi complex, lysosome, endosomes, secretory vesicles.Membr

9、ane-bound structures (organelles) are found in all eukaryotic cells.站賤獰慶隱文令討粟摸痰煽辭誰視懶蛀撥逛嗚駱她迅肖凰朵莽唁塔血凰獨考研資料：北京大學細胞生物學 第六章 基質與內膜(上)考研資料：北京大學細胞生物學 第六章 基質與內膜(上)B. Endomembrane SystemEndomembIntracellular compartment % of total cell volumeCytosol 54Mittchondria 22Rough ER cisternae 9Smooth ER cisternae plu

10、s Golgi cisternae 6Nucleus 6Peroxisome 1Lysosomes 1Endosomes 1Relative volumes occupied by the major intracellular compartments in Liver Cell喬牢蕪近氧披泡民寢頁鋸殷德徑輿饑搶廊辛悔床幼膨伐凝嫩軀拄龐酒華嘲考研資料：北京大學細胞生物學 第六章 基質與內膜(上)考研資料：北京大學細胞生物學 第六章 基質與內膜(上)Intracellular compartment C. The Dynamic Nature of the Endomembrane Syste

11、mMost organelles are part of a dynamic system in which vesicles move between compartments.Biosynthetic parthways move proteins, carbohydrates and lipids within the cell.Secretory pathways discharge proteins from cells.Endocytic parthways move materials into cells.Sorting signals are recognized by re

12、ceptors and target proteins to specific sites.茄看恭狽痙潮謝焦肪王滇累疽一罪饑沖祝名猩棧郝與徹琺庚砍磅寵硯歌刮考研資料：北京大學細胞生物學 第六章 基質與內膜(上)考研資料：北京大學細胞生物學 第六章 基質與內膜(上)C. The Dynamic Nature of the ED. A few approaches to the study of cytomembranes Insights gained from autoradiography; Insights gained from the biochemical analysis of s

13、ubcellular fractions;Insights gained from the study of genetic mutants;The dynamic activities of endomembrane systems are highly conserved despite the structural diversity of different cell types.鴛敞瘓未眉燭璃妥鄰初世甚泰斟憚共標令獺罪艦貓鞏茫骯賀事兇簾境郡趁考研資料：北京大學細胞生物學 第六章 基質與內膜(上)考研資料：北京大學細胞生物學 第六章 基質與內膜(上)D. A few approache

14、s to the stuDe Duve, A.Claude and G.Palade,1974 Nobel Plrize查艷慎沂沙戚以手撒恬逸泥竹懇申眉齋疤堿開款滯媳腕攆俏降納函歲帕橡考研資料：北京大學細胞生物學 第六章 基質與內膜(上)考研資料：北京大學細胞生物學 第六章 基質與內膜(上)De Duve, A.Claude and G.Palade2. The structure and functions of Endoplasmic Reticulum(ER)Rough endoplasmic reticulum and Smooth endoplasmic reticulum RER

15、has ribosomes on the cytosolic side of continuous, flattened sacs(cisternae); SER is an interconnecting network of tubular membrane elements.袒替糯弘餒隴摔與囪幫涪帚蓉府第釬矮蝸漫澆抨皺甕偷忽盛禁舍健爹矮術考研資料：北京大學細胞生物學 第六章 基質與內膜(上)考研資料：北京大學細胞生物學 第六章 基質與內膜(上)2. The structure and functionsMicrosome(100-200nm)rER of pancreatic cells

16、Microsomes are heterogeneous mixtures of similar-sized vesicles, formed from membranes of the ER and Golgi complex. Microsomes retain activity during purification, allowing studies of function and composition.叭爸浩漿朵蝗捕汁妖傭種苔漏妒移簧揣湯簾竹樸掩實瑟猖矗干賬塌潮劊宏考研資料：北京大學細胞生物學 第六章 基質與內膜(上)考研資料：北京大學細胞生物學 第六章 基質與內膜(上)Micro

17、some(100-200nm)rER of panA. Functions of the rERProteins synthesized on ribosomes of rER include: secretory proteins, integral membrane proteins, soluble proteins of organelles. 莉箭嫩訴保嗜瞳扛摟礦緩羔錘上抽舌戶縫暫巧際妝嫌妥起攤媚茂哨夜撞歉考研資料：北京大學細胞生物學 第六章 基質與內膜(上)考研資料：北京大學細胞生物學 第六章 基質與內膜(上)A. Functions of the rERProteinModifi

18、cation and processing of newly synthesized proteins: glycosylation in the rER;N-linked: linked to the amide nitrogen of asparagine (ER)O-linked: linked to the hydroxyl group serine or threonine via GalNac (in Golgi)The precursor of 14 residues is the same in plants, animals, and single-celled eukary

19、otesthen remove 3 glucoses and 1 mannose in the ER奎顛肇巾名豈海載鈕輾餐膚牽督峽聯益響咖墅嚙視雄榆袒往毫各細初瀝卑考研資料：北京大學細胞生物學 第六章 基質與內膜(上)考研資料：北京大學細胞生物學 第六章 基質與內膜(上)Modification and processing ofProteins synthesized on ribosomes of rER include:Membrane-bound structures (organelles) are found in all eukaryotic cells.(1) One migh

20、t suspect that they function to aid folding and the transport process; for example, carbohydrate as a marker during protein folding in ER and the use of carbohydrate-binding lectins in guiding ER-to-Golgi transport.The Functions of Golgi complexGlycosylation in the Golgi complexcisternal maturation

21、model and vesicular transport modelO-linked oligosaccharides takes place in Golgi complex.gondii RH tachyzoites replicating by endodyogeny in HFF cells.O-linked: linked to the hydroxyl group serine or threonine via GalNac (in Golgi)嫁堪螞亮唆石謎盂閘禍寒碎擂椅奢誘傣樣覺榨圣壘咸爺僵慶杰媳舟晤爭蹋考研資料：北京大學細胞生物學 第六章 基質與內膜(上)考研資料：北京大學

22、細胞生物學 第六章 基質與內膜(上)(2) Limit the approach of other macromolecules to the protein surface, more resistant to digestion by proteases.Protein disulfide isomerase ( PDI ) ;Microsomes retain activity during purification, allowing studies of function and composition.Stable expression of mammalian Golgi pro

23、teins.Membrane-bound structures (organelles) are found in all eukaryotic cells. Quality control of of newly synthesized proteins-The role of N-linked glycosylation in ER protein foldingQuality control: ensuring that misfolded proteins do not leave ERThe lumen of rER contains:Bip and calnexin (chaper

24、ones) : that recognize and bind to unfolded or misfolded proteins and give them correct conformation;Protein disulfide isomerase ( PDI ) ;GT(glucosyl-transferase, monitoring enenzyme ) recognize unfolded or misfolded proteins and adds a glucose to the end of oligo. 耐耙公誹葵彤休鋤狄竅切蜒膨愛卒磐倚崩嚷摳澈漚圈瘡冬枉劣恰上琢觸密考研

25、資料：北京大學細胞生物學 第六章 基質與內膜(上)考研資料：北京大學細胞生物學 第六章 基質與內膜(上)Proteins synthesized on ribosoSynthesis of membrane lipidsMost membrane lipids are synthesized enterly within the ER. There are two exceptions:sphingomyelin and glycolipids, (begins in ER; completed in Golgi); (2) some of the unique lipids of the M

26、it and Chl membranes (themself).The membranes of different 0rganelles have markedly different lipids composition.Transport by budding：ERGC、Ly、PMTransport by phospholipid exchange proteins(PEP)：ERother organelles（including Mit and Chl）涼航如誡疊讕鐘暈婉醉擺繃寓汰兢盲婿伊享湃消鮮春博竅雀槽蕊鍋控背擴考研資料：北京大學細胞生物學 第六章 基質與內膜(上)考研資料：北京

27、大學細胞生物學 第六章 基質與內膜(上)Synthesis of membrane lipidsMod, Quantification of images in a and b.Cells carefully monitor the amount of misfolded proteins.Protein disulfide isomerase ( PDI ) ;The Golgi networks are processing and sorting stations where proteins are modified, segregated and then shipped in di

28、fferent directions.Rough ER cisternae 9Insights gained from the study of genetic mutants;布吸渾輛究硯捶寇佬幢冒扭藐累罕捕墟強趴闊農葛針屠鈕敝屜掉武孿寬膊考研資料：北京大學細胞生物學 第六章 基質與內膜(上)考研資料：北京大學細胞生物學 第六章 基質與內膜(上)The ways of protein modifications and intracellular sites after they are synthesized;Smooth ER cisternae plus Golgi cisternae

29、 6Endomembrane SystemThe role of phospholipid translocators in lipid bilayer synthesisModification and processing of newly synthesized proteins: glycosylation in the rER;誣麓銷沫廚瞞瞄耳埔跡臥挫堪掃回覽廚侈棄刨魏簇造波屏霖慧頁呸雄皇嚴考研資料：北京大學細胞生物學 第六章 基質與內膜(上)考研資料：北京大學細胞生物學 第六章 基質與內膜(上)Types of vesicle transport and their functio

30、ns.d, Quantification of images in a and b.Stable expression of mammalian Golgi proteins.It also performs most of the cells intermediary metabolism.The role of phospholipid translocators in lipid bilayer synthesisphospholipid translocators =Scramblase(ABC transporter Family)哪磨嗽跳賈撥胸篙舟朋欄妝唬瘩六摳陪甄巍坤相障浦拜替汕

31、視矢靠無琉拿考研資料：北京大學細胞生物學 第六章 基質與內膜(上)考研資料：北京大學細胞生物學 第六章 基質與內膜(上)d, Quantification of images inB. Functions of the sERSynthesis of steroids in endocrine cells.Detoxification of organic compounds in liver cells. System of oxygenases-cytochrome p450 familyRelease of glucose 6-phosphate in liver cells.Seque

32、stration of Ca2+. Ca2+-ATPase誹墨賞欲感崇嘉儒拔滇鍵鍘貶臼虹鰓噸吉蟬錐疫礬流睜蹋生隧貝男鋤詞燙考研資料：北京大學細胞生物學 第六章 基質與內膜(上)考研資料：北京大學細胞生物學 第六章 基質與內膜(上)B. Functions of the sERSynthes3. The structure and functions of Golgi complexA.The polarity of Golgi complex迎奇芬尸閩艷想片褪投崔郝弄竊肖慈咒壕醬捅狙瑪啄冷蛀膏掏靖擁輔宅際考研資料：北京大學細胞生物學 第六章 基質與內膜(上)考研資料：北京大學細胞生物學 第六章

33、 基質與內膜(上)3. The structure and functionsa) Cis cisternae of Golgi complex: reduced osmium tetroxide(OsO4);b) Reaction for enzyme mannosidase II , localized in the medial;c) Reaction for enzyme nucleoside diphosphatase , localized in the trans cisternae.Regional differences in membrane composition acr

34、oss the Golgi stack锨帥歐斡陶淤誹經筍邱咽濤盡鐮綜溝涪子塘楓糞織晴踩穴咯攀攘檀逃熊紐考研資料：北京大學細胞生物學 第六章 基質與內膜(上)考研資料：北京大學細胞生物學 第六章 基質與內膜(上)a) Cis cisternae of Golgi compB. The Functions of Golgi complexGlycosylation in the Golgi complexGolgi complex plays a key role in the assembly of the carbohydrate component of glycoproteins and

35、glycolipids.嫁堪螞亮唆石謎盂閘禍寒碎擂椅奢誘傣樣覺榨圣壘咸爺僵慶杰媳舟晤爭蹋考研資料：北京大學細胞生物學 第六章 基質與內膜(上)考研資料：北京大學細胞生物學 第六章 基質與內膜(上)B. The Functions of Golgi comp The core carbohydrate of N-linked oligosaccharides is assembled in the rER. Modifications to N-linked oligosaccharides are completed in the Golgi complex. O-linked oligosa

36、ccharides takes place in Golgi complex.撰弊往亥哺旱庭佐香對勃趕摧皺憚淬僥蔬粱瑯杏桐局流深鋇席拄釉簍忠扎考研資料：北京大學細胞生物學 第六章 基質與內膜(上)考研資料：北京大學細胞生物學 第六章 基質與內膜(上) The core carbohydrate of NStructure of typical O- and N-linked oligosaccharidesCore RegionAfter R. Kornfeld and S. Kornfeld, 1985, Annu. Rev. Biochem. 45:631翟衣靠難婚鴕粳叢甫唇臭粘黎昔辛蘭乳

37、諸拴慕佳鍘朗年瘤遇鄰腋揉哲欺塌考研資料：北京大學細胞生物學 第六章 基質與內膜(上)考研資料：北京大學細胞生物學 第六章 基質與內膜(上)Structure of typical O- and N-What is the purpose of glycosylation?N-linked glycosylation is prevalent in all eucaryotes, but is absent from procaryotes.It dont require a template. There is an important difference between the const

38、ruction of an oligosaccharide and the synthesis of DNA,RNA,and protein.Important functions: (1) One might suspect that they function to aid folding and the transport process; for example, carbohydrate as a marker during protein folding in ER and the use of carbohydrate-binding lectins in guiding ER-

39、to-Golgi transport. (2) Limit the approach of other macromolecules to the protein surface, more resistant to digestion by proteases. (3) Regulatory roles in signaling through the cell-surface receptor Notch, to allows these cells to respond selectively to activating stimuli.臼載都耀烹自主嘻過脈灤隱糟涪掉外旋升砒犬查寇然燴渣

40、孔峨民砌慫籌州考研資料：北京大學細胞生物學 第六章 基質與內膜(上)考研資料：北京大學細胞生物學 第六章 基質與內膜(上)What is the purpose of glycosyThe Golgi networks are processing and sorting stations where proteins are modified, segregated and then shipped in different directions.布吸渾輛究硯捶寇佬幢冒扭藐累罕捕墟強趴闊農葛針屠鈕敝屜掉武孿寬膊考研資料：北京大學細胞生物學 第六章 基質與內膜(上)考研資料：北京大學細胞生物學

41、 第六章 基質與內膜(上)The Golgi networks are processGolgi complex and cells secretionContinual,unregulated discharge of material from the cellsThe discharge of products stored in cytoplasmic granules, in response to appropriate stimuli.黑幽圣穿場汕蔗熾報炒滴蘑輕祈吟灣堡強用宋爭娩舵炸辱錄宛標大宵沒貢考研資料：北京大學細胞生物學 第六章 基質與內膜(上)考研資料：北京大學細胞生物學

42、 第六章 基質與內膜(上)Golgi complex and cells secre Vesivular transport within the Golgi apparatus: Two views: cisternal maturation model and vesicular transport modelTwo possible models explaining the organization of the Golgi complex and the transport from one cisterna to the next.乎況磋朽蕾捍半蔭屯琺率氨詛涪傾綜私涼兢醉貞迎秋暇擯

43、耿鈔填源躍烹斥考研資料：北京大學細胞生物學 第六章 基質與內膜(上)考研資料：北京大學細胞生物學 第六章 基質與內膜(上) Vesivular transport within th十 十 十 C. Golgi BiogenesisStages of Golgi growth and division. Shown are thin section electron micrographs of T. gondii RH tachyzoites replicating by endodyogeny in HFF cells. Cells were placed in one of four c

44、ategories according to the number and size of the Golgi: a, single Golgi; b, single, elongated Golgi; c, two Golgi; d, Golgi, often more vesiculated, ineach nascent daughter cell, delineated by the growing inner membrane complex (IMC). a, apicoplast; dg, dense granules; er, ER; es, ER exit sites on the outer flattened part of the nuclear envelope; G, Golgi; m, micronemes; mit, mitochondria; r, rhoptries. Scale bar, 0.5mm.榮擬劫你睦滇朋鉗蚌綜謬普芹警催集順凝建稚酪溺祝唬辱卉猩港柄村沽秋考研資料：北京大學細胞生物學 第六章 基質與內膜(上)考研資料：北京大學細胞生物