세포 및 분자생물학 특론

연세대학교
Jürgen Roth

페이스북 트위터 카카오톡 네이버밴드 링크복사

주제분류

자연과학 >생물ㆍ화학ㆍ환경 >생명공학
강의학기

2013년 1학기

조회수

5,328

The structure and function of exocytosis (secretion) as part of the late secretory pathway and of endocytosis, which is closely coupled to it, will be presented at the molecular, supramolecular and organelle level together with aspects related to their genomics, proteomics and glycomics

수강안내 및 수강신청
※ 수강확인증 발급을 위해서는 수강신청이 필요합니다

Formation and dynamics of post-Golgi apparatus carriers. Constitutive secretion. Sorting in the TGN: membrane proteins and lipids.

차시별 강의

1.	Formation and dynamics of post-Golgi apparatus carriers. Constitutive secretion. Sorting in the TGN: membrane proteins and lipids.	Epithelial versus mesenchymal cells. Junctional complexes and cell polarity. The TGN as a sorting station: morphology and function. TGN marker proteins. Different transport carriers originate from the TGN. Molecular biology of clathrin coat. Adaptor proteins for clathrin coated vesicles. Dynamin and the fission of clathrin-coated vesicles. Uncoating of clathrin-coated vesicels.
	Formation and dynamics of post-Golgi apparatus carriers. Constitutive secretion. Sorting in the TGN: membrane proteins and lipids.	Types of secretion. constitutive versus regulated secretion. Constitutive secretion of fibrinogen: synthesis and assembly. Constitutive secretion of fibrinogen and endoproteolytical processing. Types of post_Golgi carriers: vesicles and tubules, mode of formation and budding/fission. Sorting of membrane lipids and proteins from the TGN: apical sorting versus basolateral sorting, sorting signals. Experimental analysis of sorting.
2.	Deborde S, Perret E, Gravotta D, Deora A, Salvarezza S, Schreiner R, Rodriguez-Boulan E (2008) Clathrin is a key regulator of basolateral polarity. Nature 452:719-723	Presentation and discusssion of a scientific publication on the regulation of insulin secretion by clathrin.
	Formation of carriers for regulated exocytosis. Types of secretory granules and storage. Proteolytic processing and packing of pro-proteins: biology and pathology.	Regulated exocytosis of membrane proteins: gastric H-ATPase and hydrochloric acid secretion. Regulated exocrine versus endocrine secretion, paracrine secretion of hormones. Types and structure of exocrine and endocrine secretory granules. Synaptic vesicles. Formation of secretory vesicles: formal steps and cell biology aspects. Sorting signals for secretory proteins and peptides in immature secretory granules. Sorting and endoproteolytic processing of pro.proteins. Family of endoproteases: phylogeny and mode of action. Proinsulin-insulin conversion and secretory granule formation. Sorting of secretory proteins and peptides by aggregation. Formation of melanosomes. General principles of pro-protein processing and sorting or endoproteases.
3.	Regulated secretion: mechanism of stimulation, interaction between secretory granule and plasma membrane.	Intracellular movement, distribution and storage of mature secretory granules. Stimuli for regulated secretion. Cellular pools of secretory granules. Time course and selectivity of regulated exocytosis. Insulin secretion. Targeting of secretor granules by Rab proteins. Fusion of secretory granules by SNARE proteins. Molecular biology of Rab and SNARE proteins and interaction with SM proteins. SNARE conformational cycle for regulated secretion. Exocytosis of synaptic vesicles. Integrated OMICS of synaptic vesicles.
4.	Engel S, Heger T, Mancini R, Herzog F, Kartenbeck J, Hayer A, Helenius A. The role of endosomes in SV40 entry and infection. J Virol 85: 4198-42112011	Presentation and discussion of a publication on virus entry in cells and the role of caveolae.
	Endocytosis: receptor-mediated endocytosis via clathrin-coated pits. Molecular biology of clathrin coats. Caveolar and non-caveolar endocytosis. Molecular biology and cell type-specificity of caveolins.	Types of clathrin-coated structures and location. Structure of clathrin-coated pits and clathrin-coated vesicles. Molecular structure of clathrin and adaptor proteins. Mode of formation of clathrin-coated pits Mode of fission of clathrin-coated vesicles. Synaptic vesicle retrieval by compensatory membrane internalization. Structure, composition and function of clathrin-coated plaques and comparison with clathrin-coated pits.
5.	Guerra L, Teter K, Lilley BN, Stenerlow B, Holmes RK, Ploegh HL, Sandvig K, Thelestam M, Frisan T (2005) Cellular internalization of cytolethal distending toxin: a new end to a known pathway. Cell Microbiol 7:921-934	Presentation and discussion of a publication on entry of a toxin in cells and its mechanism of action.
	Endosome formation and types of endosomes. Endocytotic pathways taken by pathogens and toxins.	Different intracellular sites are reached by clathrin-coated vesicles and by caveolae. Molecular characteristics of early, late and recycling endosomes. Immunocytochemistry of the endo-lysosomal system. Molecular cell biology of receptior recycling and cell type-speciific recycling endosomes, pericentriolar tubular endosomes. Recycling endosomes and glucose transporter. Birbeck granules of skin Langerhans cells and antigen presentation. Structure, formation and function of multivesicular bodies.
6.	Eden ER, White IJ, Tsapara A, Futter CE (2010) Membrane contacts between endosomes and ER provide sites for PTP1B-epidermal growth factor receptor interaction. Nat Cell Biol 12:267-272	Presentation and discussion of a publication about the endocytic route of the EGF receptor.
	Transcytosis in epithelial cells and endothelial cells. Endocytic routes and the fate of endocytosed macromolecules and receptors.	Transcytosis occurs by epithelial cells. Function of transcytosis on body defense against pathogens. Dimeric IgA, pentameric IgM, and monomeric IgG for mucosal immunity. Mode of production and of cellular passage of dimeric IgGA. The polymeric immunoglobulin receptor in basolateral IgGA uptake. Mode of action of the polymeric immunoglobulin receptor and its processing for apical secretion. Transcytosis of IgG in humans and rodents by the FcRn. Morphology of transcytosis carriers.
7.	Shen B, Wu N, Yang J-M, Gould SJ (2011) Protein Targeting to Exosomes/ Microvesicles by Plasma Membrane Anchors.	Presentation and discussion of a scientific publication about the targeting of membrane proteins to the exosome for subsequent secretion.
	Endocytic routes and the fate of endocytosed macromolecules and receptors. The exosome	The route taken by the LDL receptor and its fate by recycling. The mechanism and functional consequences of EGF binding to its receptor and the fate of the ligand-receptor complex. Insulin to insulin-receptor binding, functional consequences and lysosomal digestion of ligand-receptor complex. The components of the retromer and its function in receptor recycling. The WASH complex for tubule fission from recycling endosomes. Cellular functions of the retromer.
8.	Phagocytosis: cellular aspects and molecular mechanism Physiological functions and pathology	Synopsis of endocytic routes and comparison with phagocytosis. Evolutionary aspects of phagocytosis and its role for nutrition acquisition, host defense, embryogenesis and tissue remodeling. Mechanism of phagocytosis and cellular as well as tissue aspects. Definition and function of tissue-specific macrophages and selected examples of their physiological role for uptake of cells and pathogens. Phagocytosis and red cell maturation and aging. Phagocytosis and removal of apoptotic bodies. Clearance of rod receptor discs by phagocytosis through the retinal pigment epithelium. Alveolar macrophages in the clearance of environmental particular pollutants
9.	van der Wel N, et al. (2007) M. tuberculosis and M. leprae translocate from the phagolysosome to the cytosol in myeloid cells. Cell 129:1287-1298	Presentation and discussion of a scientific publication about translocation of pathogenic bacteria by phagocytes.
	Phagocytosis: killing of bacteria. Peroxisomes: Origin, structure, functional subcompartments, cell type specificity. Biogenesis of peroxisomes.	Phagocytosis mechanism of bacteria by phagocytes e.g. neutrophilic leukocytes. Importance of ROS, for bacterila killing in phagocytes. The role of NADPH oxidase in generation of ROS. Generation of ROS by myeloperoxidase of azurophilic granules of neutrophilic leukocytes. Formation of hypochloric acid by NOS. ROS induced damage of DNA, protein and lipids, and lysozyme-mediated digestion of bacterial capsule. Escape routes of various pathogenic bacteria from phgocytosis-mediated killing. Synopsis of TGN carriers, exocytosis and the various endocytosis pathways.
	Phagocytosis: killing of bacteria. Peroxisomes: Origin, structure, functional subcompartments, cell type specificity. Biogenesis of peroxisomes.	General introduction to peroxisomes and mitochondria as oxygen-using organelles and their function in cellular metabolism. Critical evaluation of the origin of peroxisomes: endosymbiosis versus endogenous organelle. Fine structure of peroxisomes and microscopic techniques to visualize and study peroxisomes. Cell type-specific variations in size and shape and the presence of peroxisomal networks. Importance of microtubules for peroxisomal positioning and mobility. Compartmentalization of peroxisomal oxidative enzymes and their induction by pollutants or drugs and xenobiotika. Mechanisms of biogenesis of peroxisomes: de novo formation versus division and growth model. Classification of peroxisomal membrane proteins, their receptor proteins and modes of biosynthesis on free polysomes. Synthesis and membrane insertion of peroxisomal phospholipids. The types and structure of peroxisomal matrix proteins and their functions.
10.	Munck JM, Motley AM, Nuttall JM, Hettema EH (2009) A dual function for Pex3p in peroxisome formation and inheritance. J Cell Biol 187:463-471	Presentation and discussion of a scientific publication on the mechanism of de nove formation of peroxisomes from the ER
	Peroxisomes: biogenesis and disorders Importance for cellular metabolism Free radical generation and elimination	Peroxisomes have no own genome and all their proteins are made by the host genome on free polysomes. The role of Pex3 and Pex19 in the de novo biogenesis of pre-peroxisomes from the endoplasmic reticulum. Maturation of pre-peroxisomes by growth through post-translational membrane protein insertion. Internal membrane targeting sequences of peroxisomal membrane proteins for binding to and insertion into the membrane. Synthesis and mode of membrane isnertion of phospholipids for peroxisomes. Biosynthesis of peroxisomal matrix proteins and the different types of targeting signals (PST1 and PST2). The molecular composition and function of the peroxisomal matrix protein importomer. The importomer cycle. The multiplication of peroxisomes by elongation through Pex11p, constriction and fission through DLP1 and Fis1.
	Peroxisomes: biogenesis and disorders Importance for cellular metabolism Free radical generation and elimination	Function of peroxisomes in beta oxidation of fatty acids, plasmalogen biosynthesis, glyoxylate and amino acid metabolism, polyamine oxidation, cholesterol metabolism, and beta oxidation of leukotrienes and prostaglandins. Function of peroxisomal oxidases in the generation oxygen-derived free radicals and the detoxification of metobolic intermediates. The involvement of peroxisomes in the aging process. Peroxisomal disorders such as deficiencies of single enzymes or biogenesis disorders. Mice knock out models of peroxisomal disorders.
11.	Grewal et al.: The Ashwell receptor mitigates the lethal coagulopathy of sepsis. Nature Med 14: 648, 2008	Presentation and discussion of a scientific publication on the importance of the Ashwell receptor for the coagulation system during sepsis
	Mitochondria: structure and subcompartments Biosynthesis and targeting of membrane and matrix proteins Importance for cellular metabolism and apoptosis Diseases due to mitochondrial abnormalities	Fine structure of mitochondria and differences to peroxisomes. The mitochondrial compartiments and their general functions. Fine structure and mitochondrial networks. Cristae and tubuli-type of mitochondria and their tissue-specific distribution. Molecular basis for the formation of the cristae junction and cristae tips. Endoplasmic reticulum-mitochondria complexes: molecular composition and function. The role of mitofusin 2 and of trichoplein for ER-mitochondria contacts. Multiplication of mitochondria occurs by fission and the involvement of Dnm, Mdv1 and Fis1 proteins. The mitochondrial fission machinery of yeast and vertebrates. Mitochondrial dynamics involve fission and fusion and the formation of networks as well as removal by mitophagy.
	Mitochondria: structure and subcompartments Biosynthesis and targeting of membrane and matrix proteins Importance for cellular metabolism and apoptosis Diseases due to mitochondrial abnormalities	The mitochondrial fusion machinery of yeast and vertebrates. The diverse functions of the different fusion machinery proteins in the cellular metabolism. The importance of OPA1 for mitochondrial function and the initiation of apoptosis. The life cycle of mitochondria related to the cell cycle. The mitochondrial genome and the types of proteins rRNA and tRNA it is coding for. A transcriptional network controls mitochondrial gene expression. The different classes of outer and inner mitochondrial membrane proteins.