title: A residue in the S2 subsite controls substrate selectivity of matrix metalloproteinase-2 and matrix metalloproteinase-9.
abstract: Matrix metalloproteinase (MMP)-2 and MMP-9 are closely related metalloproteinases that are implicated in angiogenesis. The two proteins have a similar domain structure and highly homologous catalytic domains, making them an excellent comparative model for understanding the structural basis of substrate recognition by the MMP family. Although the two MMPs exhibit some overlap in substrate recognition, our recent work showed that MMP-2 can cleave a set of peptide substrates that are only poorly recognized by MMP-9 (Chen, E. I., Kridel, S. J., Howard, E. W., Li, W., Godzik, A., and Smith, J. W. (2002) J. Biol. Chem. 277, 4485-4491). Mutations at the P(2) position of these peptide substrates dramatically reduced their selectivity for MMP-2. Inspection of the corresponding S(2) pocket of the substrate-binding cleft of the protease reveals that MMP-9 contains an Asp, whereas MMP-2 contains Glu. Here, we test the hypothesis that this conservative substitution has a role in substrate selectivity. Mutation of Glu(412) in MMP-2 to Asp significantly reduced the hydrolysis of selective substrates, with only a minor effect on hydrolysis of non-selective substrates. The predominant effect of the mutation is at the level of k(cat), or turnover rate, with reductions reaching as high as 37-fold. The residues that occupy this position in other MMPs are highly variable, providing a potential structural basis for substrate recognition across the MMP family.
authors.affiliation: Cancer Research Center, The Burnham Institute, La Jolla, CA 92037, USA.
authors.forename: Emily I
authors.initials: EI
authors.lastname: Chen
authors.forename: Weizhong
authors.initials: W
authors.lastname: Li
authors.forename: Adam
authors.initials: A
authors.lastname: Godzik
authors.forename: Eric W
authors.initials: EW
authors.lastname: Howard
authors.forename: Jeffrey W
authors.initials: JW
authors.lastname: Smith
mesh_terms: D019943:Amino Acid Substitution; D002460:Cell Line; D006801:Humans; D006868:Hydrolysis; D020778:Matrix Metalloproteinase 2 / Q000235:genetics / Q000378:metabolism*; D020780:Matrix Metalloproteinase 9 / Q000235:genetics / Q000378:metabolism*; D009154:Mutation; D013379:Substrate Specificity / Q000235:genetics
chemical_list: D020778:Matrix Metalloproteinase 2; D020780:Matrix Metalloproteinase 9