title: Coupling protein engineering with probe design to inhibit and image matrix metalloproteinases with controlled specificity.
abstract: Matrix metalloproteinases (MMPs) are zinc endopeptidases that play roles in numerous pathophysiological processes and therefore are promising drug targets. However, the large size of this family and a lack of highly selective compounds that can be used for imaging or inhibition of specific MMPs members has limited efforts to better define their biological function. Here we describe a protein engineering strategy coupled with small-molecule probe design to selectively target individual members of the MMP family. Specifically, we introduce a cysteine residue near the active-site of a selected protease that does not alter its overall activity or function but allows direct covalent modification by a small-molecule probe containing a reactive electrophile. This specific engineered interaction between the probe and the target protease provides a means to both image and inhibit the modified protease with absolute specificity. Here we demonstrate the feasibility of the approach for two distinct MMP proteases, MMP-12 and MT1-MMP (or MMP-14).
authors.affiliation: Department of Pathology, Stanford University School of Medicine, Stanford, California 94305, USA.
authors.forename: Montse
authors.initials: M
authors.lastname: Morell
authors.forename: Thinh
authors.initials: T
authors.lastname: Nguyen Duc
authors.forename: Amanda L
authors.initials: AL
authors.lastname: Willis
authors.forename: Salahuddin
authors.initials: S
authors.lastname: Syed
authors.forename: Jiyoun
authors.initials: J
authors.lastname: Lee
authors.forename: Edgar
authors.initials: E
authors.lastname: Deu
authors.forename: Yang
authors.initials: Y
authors.lastname: Deng
authors.forename: Junpeng
authors.initials: J
authors.lastname: Xiao
authors.forename: Benjamin E
authors.initials: BE
authors.lastname: Turk
authors.forename: Jason R
authors.initials: JR
authors.lastname: Jessen
authors.forename: Stephen J
authors.initials: SJ
authors.lastname: Weiss
authors.forename: Matthew
authors.initials: M
authors.lastname: Bogyo
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D003545:Cysteine / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D006801:Humans; D020781:Matrix Metalloproteinase 1 / Q000032:analysis* / Q000235:genetics / Q000378:metabolism; D053508:Matrix Metalloproteinase 12 / Q000032:analysis* / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008958:Models, Molecular; D015336:Molecular Probe Techniques; D008969:Molecular Sequence Data; D061848:Optical Imaging; D015202:Protein Engineering / Q000379:methods*; D016415:Sequence Alignment; D015027:Zebrafish
chemical_list: D053508:Matrix Metalloproteinase 12; D020781:Matrix Metalloproteinase 1; D003545:Cysteine