MATRIX METALLOPROTEINASE Andressa Obice Tony Voong November 29

MATRIX METALLOPROTEINASE Andressa Obice &Tony Voong November 29, 2010 Biochemistry

Overview Family of highly-homologous, zinc dependent endopeptidases Groups: collagenases, gelatinases, stromelysins, matrilysins, membrane-type MMPs (MT-MMPs), and Other MMPs Function: -degrade matrix/ non-matrix proteins Biological processes: -embryogenesis, tissue repair/remodeling, wound healing, angiogenesis Diseases: -atheroma, arthritis, cancer, tissue ulceration, nephritis, fibrosis, aneurysms Regulated by tissue inhibitors of metalloproteinases (TIMPS)

Structure Most MMPs: -propetide domain -catalytic domain -hinge region -hemopexin domain MMP-7, MMP-26, MMP-23, - no hinge region - no hemopexin domain MMP-23 -cysteine-rich domain, -immunoglobulin-like domain

Propeptide domain Catalytic domain Hinge region About 80 amino acids • About 170 amino acids • Variable lengths • No determinable structure Must be removed before • Spherical shape • Catalytic zinc ion enzyme is active (zinc-binding motif) Hemopoxin Domain “Cysteine switch” • “Met-turn” • About 200 amino acids Furin cleavage site • Fibronectin type II repeats

Tertiary (3 D) Structure of MMPs Catalytic Domain: 5 -stranded-β-pleated sheet 3 α-helices Connective loops 2 zinc ions (1 catalytic/1 structural) Up to 3 calcium ions 3 histidines “S 1’ pocket” Fibronectin Type II domains -2 antiparrallel β sheets, connected with a short α-helix, 2 disulfide bonds

Propetide Domain 3 α chains connecting loops Hemopoxin Domain 4 -bladed β-propeller 1 disulfide bond (first/fourth blades) 1 calcium ion 1 chloride ion

Propeptide Domain Catalytic Domain Fibronectin Domain Hemopexin Domain Pro. MMP-2 -TIMP-2 Complex

Inhibition of MMP by TIMP

Function

Degradation ECM Over expression of MMP Alterations of TIMP Proteolytic activity

Therapeutics Pro TNF- TNF - converting enzyme (TACE) MMP Rheumatoid arthritis Crohn’s disease Cancer Stroke

TNF- Pro TNF- s. TNF- Stages of Inhibition 1. pro-TNF- processing 2. pro-TNF- synthesis

TACE Catalytic Domain

TACE and MMP Inhibition of TACE and MMP-1

Channel

TACE Inhibitor

TMI-1 & TAPI-2 Vs. TACE & ADAM 10 Future…

Questions?

References Chen, Taosheng. A Practical Guide to Assay Development and High-throughput Screening in Drug Discovery. Boca Raton: CRC, 2010. Print. 26 "Crystal Structure of the Catalytic Domain of Human Tumor Necrosis Factor-α-converting Enzyme — PNAS. " Proceedings of the National Academy of Sciences. Web. 5 Nov. 2010. Das. Gupta, Sirshendu, et al. , Current perspective of TACE inhibitors: A review, Bioorganic & Medical Chemistry. (2008), doi: 10. 1016/j. bmc. 2008. 11. 067 E. F. Healy, et al. , Acetylenic inhibitors of ADAM 10 and ADAM 17: In silico analysis of potency and selectivity, J. Mol. Graph. Model. (2010), doi: 10. 1016/j. jmgm. 2010. 08. 006 Levin, J. I. , et al. Acetylenic TACE inhibitors. Part 2: SAR of six-membered cyclic sulfonamide hyroxamates, Bioorganic & Medical Chemistry. (2005), doi: 10. 1016/j. bmcl. 2005. 06. 072 M. J. Eck, S. R. Sprang. The Structure Of Tumor Necrosis Factor-Alpha At 2. 6 A Resolution. Implications For Receptor Binding. J. Biol. Chem. V. 264 17595 1989 Rao, B. Govinda, et al. , Novel thiol-based TACE inhibitors: Rational design, synthesis, and SAR of thiol containing aryl sulfonamides, Bioorganic & Medical Chemistry. (2007), doi: 10. 1016 j. bmcl. 2007. 01. 064