title: Additivity-based design of the strongest possible turkey ovomucoid third domain inhibitors for porcine pancreatic elastase (PPE) and Streptomyces griseus protease B (SGPB).
abstract: We describe here successful designs of strong inhibitors for porcine pancreatic elastase (PPE) and Streptomyces griseus protease B (SGPB). For each enzyme two inhibitor variants were designed. In one, the reactive site residue (position 18) was retained and the best residues were substituted at contact positions 13, 14, and 15. In the other variant the best residues were substituted at all contact positions except the reactive site where a Gly was substituted. The four designed variants were: for PPE, T(13)E(14)Y(15)-OMTKY3 and T(13)E(14)Y(15)G(18)M(21)P(32)V(36)-OMTKY3, and for SGPB, S(13)D(14)Y(15)-OMTKY3 and S(13)D(14)Y(15)G(18)I(19)K(21)-OMTKY3. The free energies of association (ΔG(0)) of expressed variants have been measured with the proteases for which they were designed as well as with five other serine proteases and the results are discussed.
authors.affiliation: Department of Chemistry, Purdue University, 1393 Brown Building, West Lafayette, IN 47907-1393, USA. qasimm@ipfw.edu
authors.forename: Mohammad A
authors.initials: MA
authors.lastname: Qasim
authors.forename: Lixia
authors.initials: L
authors.lastname: Wang
authors.forename: Sabiha
authors.initials: S
authors.lastname: Qasim
authors.forename: Stephen
authors.initials: S
authors.lastname: Lu
authors.forename: Wuyuan
authors.initials: W
authors.lastname: Lu
authors.forename: Richard
authors.initials: R
authors.lastname: Wynn
authors.forename: Zheng-Ping
authors.initials: ZP
authors.lastname: Yi
authors.forename: Michael
authors.initials: M
authors.lastname: Laskowski
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D018360:Crystallography, X-Ray; D015195:Drug Design; D006801:Humans; D007700:Kinetics; D008969:Molecular Sequence Data; D010196:Pancreatic Elastase / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000235:genetics; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D012697:Serine Endopeptidases / Q000737:chemistry* / Q000235:genetics; D015842:Serine Proteinase Inhibitors / Q000737:chemistry* / Q000378:metabolism; D013305:Streptomyces griseus / Q000737:chemistry* / Q000201:enzymology; D013329:Structure-Activity Relationship; D013552:Swine; D013816:Thermodynamics; D014359:Trypsin Inhibitor, Kazal Pancreatic / Q000737:chemistry* / Q000235:genetics
chemical_list: D015842:Serine Proteinase Inhibitors; C039345:ovomucoid inhibitors; D014359:Trypsin Inhibitor, Kazal Pancreatic; D012697:Serine Endopeptidases; D010196:Pancreatic Elastase; C024984:streptogrisin B
keywords: Additivity; CARL; HLE; Inhibitor design; Kazal inhibitor; OMTKY3; PPE; Protease inhibitor; SGPA and SGPB; SRA; Serine protease; Streptomyces griseus protease A and B. In MEROPS database and recent literature these are listed as Streptogrisin A and B; human leukocyte elastase; porcine pancreatic elastase; sequence to reactivity algorithm; subtilisin Carlsberg; turkey ovomucoid third domain
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