title: Binding pockets on the surface of human leukocyte elastase and human leukocyte cathepsin G. Implications to the design of inhibitors derived from human C-reactive protein.
abstract: Analogs of the peptide Val-Thr-Val-Ala-Pro-Val-His-Ile, derived from the primary sequence of the acute phase reactant CRP, i.e. amino acid residues 89-96, were optimized to inhibit the enzymatic activities of human leukocyte elastase (hLE) and human leukocyte cathepsin G (hCG), which are associated with tissue damage occurring in the course of several chronic inflammatory conditions. hLE's major S1 pocket, lined mostly by hydrophobic amino acid residues, was shown by theoretical electrostatic potential calculations to possess some negative charge. This pocket was found to be extremely sensitive towards modifications in the P1 position of CRP derived inhibitors, with valine being the preferred amino acid. In contrast, the corresponding S1 pocket of hCG is large and accepts the positively charged 'aromatic' side chain of histidine, which increases most significantly the capability of CRP derived inhibitors. A prominent positive pocket was observed in the distant S7 region of hLE, which is generated by two exposed positive residues, Arg177 and Arg217, on the enzymes surface. This long range subsite was utilized to increase the hLE inhibitory activity of CRP derived peptide using the natural sequence of CRP, which contains a unique glutamic acid moiety in the P7 position. In contrast to the charged nature of hLE's S7 pocket, the corresponding pocket on the surface of hCG appears to be less prominent. Additional hydrophobic N-terminus modifications of CRP89-96 increased the inhibitory activity towards both enzymes, provided that residues P1 and p7, were designed according to the individual preferences of hLE and hCG. The unique interaction between the negative amino acid side chain of CRP with the positive S7 pocket of hLE as elucidated in this study, and additional subsite preferences may now be used in the design of novel therapeutic substances.
authors.affiliation: Department of Organic Chemistry, Weizmann Institute of Science, Rehovot, Israel.
authors.forename: E J
authors.initials: EJ
authors.lastname: Yavin
authors.forename: M
authors.initials: M
authors.lastname: Eisenstein
authors.forename: M
authors.initials: M
authors.lastname: Fridkin
mesh_terms: D000595:Amino Acid Sequence; D001665:Binding Sites; D002097:C-Reactive Protein / Q000737:chemistry*; D056649:Cathepsin G; D002403:Cathepsins / Q000037:antagonists & inhibitors* / Q000737:chemistry* / Q000378:metabolism; D015195:Drug Design; D004791:Enzyme Inhibitors / Q000138:chemical synthesis / Q000737:chemistry* / Q000494:pharmacology; D006801:Humans; D066298:In Vitro Techniques; D019272:Leukocyte Elastase / Q000037:antagonists & inhibitors* / Q000737:chemistry* / Q000378:metabolism; D007962:Leukocytes / Q000201:enzymology; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology; D010446:Peptide Fragments / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology; D011487:Protein Conformation; D012697:Serine Endopeptidases; D013329:Structure-Activity Relationship
chemical_list: D004791:Enzyme Inhibitors; D009842:Oligopeptides; D010446:Peptide Fragments; D002097:C-Reactive Protein; D002403:Cathepsins; D012697:Serine Endopeptidases; C534094:CTSG protein, human; D056649:Cathepsin G; D019272:Leukocyte Elastase