Lecture 7-8.pdf - MCDB 145/CHEM 151 Lecture 7 Finish...

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MCDB 145/CHEM 151 Lecture 7 Finish Collagen Polyprotein theme Essay outline due Thursday 1. Your name 2. Your topic 3. Question under investigation 4. Two major references Quiz - open-book & take-home 4 questions about variant origins Issued Thursday; due Tuesday
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Collagen in context TROPOCOLLAGEN ”squashed” microfibril In negative staining, heavy metal falls into these interstices and then scatters electrons
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• Assembly into lateral arrays last step in the Golgi • To secretory granules EXIT RER EXIT GOLGI Liquid crystals • smectic • nematic • cholesteric For order and fluidity Immature pH 7 Mature pH 5 EXIT CELL (fibroblast)
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LC: Segment-long-spacing Constance E. Brinckerhoff and Lynn M. Matrisian Nature Reviews Molecular Cell Biology 3, 207-214 (March 2002) Smectic LCs
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N- & C-procollagen peptidases - metalloproteases Quarter stagger assembly - 0.6/0.4 banding pattern 8 Propeptide removal pH 7.5 •Fibrillogenesis 10 Cross-linking •Cross-link maturation LysOx extracellular extracellular extracellular
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Formation of 1/4 stagger Smectic array fibroblast fibroblast strain-induced alignment quarter stagger fibroblasts are pulling crystallites In shear
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Vader, D., Kabla, A., Weitz, D., Mahadevan. L. (2009). Strain induced alignment in collagen gels. PLOS One 4, e5902 Harris, A. K., Stopak, D., & Wild, P. (1981). Fibroblast traction as a mechanism for collagen morphogenesis. Nature 290, 249 - 251 Reading on strain effects on collagen
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N- & C-procollagen peptidases - metalloproteases Quarter stagger assembly - 0.6/0.4 banding pattern 8 Propeptide removal pH 7.5 •Fibrillogenesis 10 Cross-linking •Cross-link maturation LysOx extracellular extracellular extracellular
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NH CH C C H 2 NH O C H 2 C H 2 C H 2 H 2 N NH CH C C H 2 NH O C H 2 C H 2 C H 2 NH 2 NH CH C C H 2 NH O C H 2 C H 2 C H O NH CH C C H 2 NH O C H 2 C H 2 C H OH NH CH C C H 2 NH O C H 2 C H 2 C H 2 H N C H 2 C H C H 2 N C H 2 C H 2 C H 2 H N Lysyl Oxidase O 2 Schiff base Collagen Cross-linking - Lysyl Oxidation chain A chain B chain A chain A chain B •• Cross-link formation H 2 O Elimination: Aldimine Hydrogenation with ageing: Lysinonorleucine Cross-link maturation Collagen Cross-linking Role of Lysine
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Covalent collagen processing Translation function Side chain chemistry pro - 3-trans-OH pro - 4-trans-OH asn - N-glyco lys - d -OH hyl - O-glyco cys -S-S- Post translation proteolysis signal N-propeptidase C-propeptidase Post secretory X-linking lys & hyl to aldehydes Schiff base to aldimines aldol condensation: 2 aldehydes reduction with aging Huge hurdles for biotech of collagen: film, beer, processed food (gummy bears, marshmallows), pharmaceutical encapsulants Emphasize hierarchical structure intracellular extracellular extracellular
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Literature - polyproteins Polyprotein processing Reading Hauser, F. et al (1991) Peptidyl a-hydroxylating mono-oxygenase from sea anemones. Biochem. Biophys. Res. Commun. 241, 509-512.
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  • Spring '09
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