title: The intrinsic protein flexibility of endogenous protease inhibitor TIMP-1 controls its binding interface and affects its function.
abstract: Protein flexibility is thought to play key roles in numerous biological processes, including antibody affinity maturation, signal transduction, and enzyme catalysis, yet only limited information is available regarding the molecular details linking protein dynamics with function. A single point mutation at the distal site of the endogenous tissue inhibitor of metalloproteinase 1 (TIMP-1) enables this clinical target protein to tightly bind and inhibit membrane type 1 matrix metalloproteinase (MT1-MMP) by increasing only the association constant. The high-resolution X-ray structure of this complex determined at 2 A could not explain the mechanism of enhanced binding and pointed to a role for protein conformational dynamics. Molecular dynamics (MD) simulations reveal that the high-affinity TIMP-1 mutants exhibit significantly reduced binding interface flexibility and more stable hydrogen bond networks. This was accompanied by a redistribution of the ensemble of substrates to favorable binding conformations that fit the enzyme catalytic site. Apparently, the decrease in backbone flexibility led to a lower entropy cost upon formation of the complex. This work quantifies the effect of a single point mutation on the protein conformational dynamics and function of TIMP-1. Here we argue that controlling the intrinsic protein dynamics of MMP endogenous inhibitors may be utilized for rationalizing the design of selective novel protein inhibitors for this class of enzymes.
authors.affiliation: Department of Structural Biology, Weizmann Institute of Science, Rehovot, Israel.
authors.forename: Moran
authors.initials: M
authors.lastname: Grossman
authors.forename: Dmitry
authors.initials: D
authors.lastname: Tworowski
authors.forename: Orly
authors.initials: O
authors.lastname: Dym
authors.forename: Meng-Huee
authors.initials: MH
authors.lastname: Lee
authors.forename: Yaakov
authors.initials: Y
authors.lastname: Levy
authors.forename: Gillian
authors.initials: G
authors.lastname: Murphy
authors.forename: Irit
authors.initials: I
authors.lastname: Sagi
mesh_terms: D018360:Crystallography, X-Ray; D004791:Enzyme Inhibitors / Q000737:chemistry; D006801:Humans; D061965:Matrix Metalloproteinase Inhibitors; D017354:Point Mutation; D011487:Protein Conformation; D017510:Protein Folding; D019715:Tissue Inhibitor of Metalloproteinase-1 / Q000737:chemistry* / Q000235:genetics
chemical_list: D004791:Enzyme Inhibitors; D061965:Matrix Metalloproteinase Inhibitors; D019715:Tissue Inhibitor of Metalloproteinase-1