1. Golgi complex BIOLOGY, Faculty of Medicine and Dentistry 2014. 10. 06. László KŐHIDAI, PhD., Assoc. Prof. Department of Genetics, Cell- and Immunobiology Semmelweis University

3. Camillo Golgi (1843-1926) Nobel prize 1906 "internal reticular apparatus"

4. Structure Saccules Saccules Tubules Tubules Vesicles Vesicles structural-functional unit: dictyosome 4-6 saccles the structure is polarized into sub-compartments the structure is polarized into sub-compartments cis Golgimedial Golgi trans Golgi cis Golgimedial Golgi trans Golgi cis Golgi network (CGN) trans Golgi network(TGN)

6. Dictyosomes of the plant cells Synthesis of complex polysaccharides of the cell wall (hemicellulose, pectins)

7. Main functions transport transport sorting sorting transformation transformation membrane wrapping membrane wrapping

8. Golgi – „Traffic manager” of the cell

9. Main pathways going in and out Golgi

12. Pulse-chase technique

13. dER Cisternaltransport Materials are forwarded together the cisterns together the cisterns Vesicular transport transport Vesicles trasport substances from cystern to cistern from cystern to cistern

14. Significance of cytoskeletal network

15. CGN the peptides arrive from the ER in vesicles the peptides arrive from the ER in vesicles they are N-glycosilated they are N-glycosilated no sorting in the ER no sorting in the ER Bidirectional transport of proteins: soluble, endogeneous proteins of the ER recycled soluble, endogeneous proteins of the ER recycled in transport vesicles - retention signal is required in transport vesicles - retention signal is required sorting and transport of lysosomal enzymes sorting and transport of lysosomal enzymes

16. Sorting and modification of lysosomal enzymes Mannose-6-phosphate (M-6-P) signaling: based on the recognition of lysosomal hydrolases based on the recognition of lysosomal hydrolases recognition of the “signal patches” (proper 3D combination recognition of the “signal patches” (proper 3D combination of amino acids) is required of amino acids) is required main working enzyme: GlcNAc-phosphotransferase main working enzyme: GlcNAc-phosphotransferase Phosphorylation of the mannoses: promotes the sorting of these enzymes promotes the sorting of these enzymes prevents the further modifications prevents the further modifications

17. N- and O- glycosylation

18. Glycosilation in the Golgi Modifications on the N-glycosilation pattern cis-Golgi: cis-Golgi: mannose-type oligosaccharides complex oligosaccharides TGN: TGN: substitution with sialic acids - negatively charged O-glycosilation: takes place mainly in the medial- and trans-Golgi takes place mainly in the medial- and trans-Golgi sidechains of Ser and Thr are glycosilated sidechains of Ser and Thr are glycosilated

19. © 2000 by Geoffrey M. Cooper

20. mannse phosphorylation (lysosomal prot.) Removal of mannose removal of mannose labelling w galactose Labelling w syalic acid Labelling w GlcNAc sorting CGN cis-Golgi medial-Golgi trans-Golgi TGN

21. Structure of M-6-P

22. Significance of M-6-P labelling

23. Other modifications glucose-amino-glycane (GAG) chains glucose-amino-glycane (GAG) chains sulphatation (proteoglycanes, Tyr res. of peptides) - TGN sulphatation (proteoglycanes, Tyr res. of peptides) - TGN proteolytic modifications - secretion vesicle proteolytic modifications - secretion vesicle

24. Synthesis of lipids in the Golgi ceramideglycolipidssphingomyelin

25. Main transport pathways from TGN

26. endosomal-lysosomal compartment endosomal-lysosomal compartment via transport vesicles - M-6-P receptors surface membrane - secretion surface membrane - secretion constitutive secretion - transports lipids and peptide components of the surface membrane and the extracellular matrix exocytosis exocytosis regulated secretion

27. acid phosphatase trans Golgi network osmium reduction cis-Golgi unstained Enzyme content of different compartments in Golgi

28. Modifications of secretory vesicles selective aggregation - TGN selective aggregation - TGN further modifications and sorting further modifications and sorting inactive precursor - active enzyme or hormone (e.g. preproinsulin - proinsulin - insulin) concentration - loss of water concentration - loss of water hydratation - e.g. proteoglygans hydratation - e.g. proteoglygans uptake some cytoplasmatic substances e.g. histamine uptake some cytoplasmatic substances e.g. histamine

29. Formation of the acrosome  Acrosome is a large lysosome and found in sperm  It contains enzymes e.g. hyaluronidase, acid phosphatase…  During the sperm differantiation several small vesicles (primary lysosomes) are formed from Golgi  thesmall vesicles are fused to form a single large lysosome - acrosome

30. Main pathway going out the Golgi

31. Alternative pathways some molecules do not synthesized on the rER some molecules do not synthesized on the rER (e.g. interleukin 1  and 1  - IL1  -IL1 , (e.g. interleukin 1  and 1  - IL1  -IL1 , basic fibroblast growth factor-bFGF) basic fibroblast growth factor-bFGF) these molecules transported by ABC-transporters these molecules transported by ABC-transporters other roles of the alternative pathway: other roles of the alternative pathway: - elimination of toxic proteins - regulation of protein concentrations in cytosol

32. Defects of sorting mechanism lysosomal enzymes do not enter the late endosomes BUT enzymes enter the constitutive secretory pathway and released I (=inclusion) cell disease: - the M-6-P signal is not formen on the enzymes - lysosomal enzymes “escape” from the cell - deficient intracellular digestion - the non-digested substances form INCLUSIONS

36. Network of membran flow in eukaryotic cells

37. Organell-dependent metabolism of lipids

38. http://www.youtube.com/watch?v=u38LjCOvDZU