1C3I

HUMAN STROMELYSIN-1 CATALYTIC DOMAIN COMPLEXED WITH RO-26-2812

Summary for 1C3I

• Entry DOI: 10.2210/pdb1c3i/pdb
• Related: 1C8T 1HFS 1SLM
• Descriptor: STROMELYSIN-1, ZINC ION, CALCIUM ION, ... (5 entities in total)
• Functional Keywords: stromelysin-1, site mutant (glu202-gln), hydrolase
• Biological source: Homo sapiens (human)
• Cellular location: Secreted, extracellular space, extracellular matrix (Probable): P08254
• Total number of polymer chains: 2
• Total formula weight: 39792.75

Authors

Steele, D.L.,Kammlott, R.U.,Crowther, R.L.,Birktoft, J.J.,Dunten, P.,Engler, J.E. (deposition date: 1999-07-27, release date: 2000-07-26, Last modification date: 2024-02-07)

Primary citation

Steele, D.L.,El-Kabbani, O.,Dunten, P.,Windsor, L.J.,Kammlott, R.U.,Crowther, R.L.,Michoud, C.,Engler, J.A.,Birktoft, J.J.
Expression, characterization and structure determination of an active site mutant (Glu202-Gln) of mini-stromelysin-1.
Protein Eng., 13:397-405, 2000

PubMed Abstract: Human stromelysin-1 is a member of the matrix metalloproteinase (MMP) family of enzymes. The active site glutamic acid of the MMPs is conserved throughout the family and plays a pivotal role in the catalytic mechanism. The structural and functional consequences of a glutamate to glutamine substitution in the active site of stromelysin-1 were investigated in this study. In contrast to the wild-type enzyme, the glutamine-substituted mutant was not active in a zymogram assay where gelatin was the substrate, was not activated by organomercurials and showed no activity against a peptide substrate. The glutamine-substituted mutant did, however, bind to TIMP-1, the tissue inhibitor of metalloproteinases, after cleavage of the propeptide with trypsin. A second construct containing the glutamine substitution but lacking the propeptide was also inactive in the proteolysis assays and capable of TIMP-1 binding. X-ray structures of the wild-type and mutant proteins complexed with the propeptide-based inhibitor Ro-26-2812 were solved and in both structures the inhibitor binds in an orientation the reverse of that of the propeptide in the pro-form of the enzyme. The inhibitor makes no specific interactions with the active site glutamate and a comparison of the wild-type and mutant structures revealed no major structural changes resulting from the glutamate to glutamine substitution.

PubMed: 10877850
DOI: 10.1093/protein/13.6.397

Experimental method

X-RAY DIFFRACTION (1.83 Å)