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Site-directed mutagenesis combined with chemical modification as a strategy for altering the specificity of the S1 and S1' pockets of subtilisin Bacillus lentus
University of Toronto, Canada.ORCID-id: 0000-0002-9577-832X
Vise andre og tillknytning
1998 (engelsk)Inngår i: Biochemistry, ISSN 0006-2960, Vol. 37, nr 17, s. 5968-5973Artikkel i tidsskrift (Fagfellevurdert) Published
Abstract [en]

By combining site-directed mutagenesis with chemical modification, we have altered the S1 and S1 pocket specificity of subtilisin Bacillus lentus (SBL) through the incorporation. of unnatural amino acid moieties, in the following manner: WT → Cys(mutant) + H3CSO2SR → Cys-SR, where R may be infinitely variable. A paradigm between extent of activity changes and surface exposure of the modified residue has emerged. Modification of M222C, a buffed residue in the S1' pocket of SBL, caused dramatic changes in k(cat)/K(M), of an up to 122-fold decrease, while modification of S166C, which is located at the bottom of the S1 pocket and is partially surface exposed, effected more modest activity changes. Introduction of a positive charge at S166C does not alter k(cat)/K(M), whereas the introduction of a negative charge results in lowered activity, possibly due to electrostatic interference with oxyanion stabilization. Activity is virtually unaltered upon modification of S156C, which is located toward the bottom of the S1 pocket and surface exposed and whose side chain is solvated. An unexpected structure-activity relationship was revealed for S166C-SR enzymes in that the pattern of activity changes observed with increasing steric size of R was not monotonic. Molecular modeling analysis was used to analyze this unprecedented structure-activity relationship and revealed that the position of the β- carbon of Cys166 modulates binding of the P1 residue of the AAPF product inhibitor.

sted, utgiver, år, opplag, sider
1998. Vol. 37, nr 17, s. 5968-5973
Emneord [en]
subtilisin, amino acid sequence, article, conformational transition, Eggerthella lenta, enzyme activity, enzyme conformation, enzyme specificity, enzyme stability, nonhuman, priority journal, site directed mutagenesis, structure activity relation, Amino Acid Substitution, Bacillus, Crystallography, X-Ray, Indicators and Reagents, Kinetics, Mass Spectrometry, Mesylates, Methionine, Models, Molecular, Mutagenesis, Site-Directed, Serine, Substrate Specificity, Subtilisins, Sulfhydryl Compounds, Titrimetry, Bacillus lentus, Bacteria (microorganisms), Felis catus
HSV kategori
Identifikatorer
URN: urn:nbn:se:kth:diva-74865DOI: 10.1021/bi9727951OAI: oai:DiVA.org:kth-74865DiVA, id: diva2:490130
Merknad

References: Bone, R., Agard, D.A., (1991) Methods Enzymol., 202, pp. 643-671; Khosla, C., Caren, R., Kao, C.M., McDaniel, R., Wang, S.-W., (1996) Biotechnol. Bioeng., 52, pp. 122-128; Sears, P., Wong, C.-H., (1996) Biotechnol. Prog., 12, pp. 423-433; Shao, P., Arnold, F.H., (1996) Curr. Opin. Struct. Biol., 6, pp. 513-518; Perona, J.J., Craik, C.S., (1995) Protein Sci., 4, pp. 337-360; Ballinger, M.D., Tom, J., Wells, J.A., (1996) Biochemistry, 35, pp. 13579-13585; Russell, A.J., Thomas, P.G., Fersht, A.R., (1987) J. Mol. Biol., 193, pp. 803-813; Wells, J.A., Cunningham, B.C., Graycar, T.P., Estell, D.A., (1987) Proc. Natl. Acad. Sci. U.S.A., 84, pp. 5167-5171; Estell, D.A., Graycar, T.P., Miller, J.V., Powers, D.B., Burnier, J.P., Ng, P.G., Wells, J.A., (1986) Science, 233, pp. 659-663; Wangikar, P.P., Rich, J.O., Clark, D.S., Dordick, J.S., (1995) Biochemistry, 34, pp. 12302-12310; Cornish, V.W., Mendel, D., Schultz, P.G., (1995) Angew. Chem., Int. Ed. Engl., 34, pp. 621-633; Kuang, H., Brown, M.L., Davies, R.R., Young, E.C., Distefano, M.D., (1996) J. Am. Chem. Soc., 118, pp. 10702-10706; Peterson, E.B., Hilvert, D., (1995) Biochemistry, 34, pp. 6616-6620; Suckling, C.J., Zhu, L.-M., (1993) Bioorg. Med. Chem. Lett., 3 (4), pp. 531-534; Rokitas, S.E., Kaiser, E.T., (1986) J. Am. Chem. Soc., 108, pp. 4984-4987; Kukubo, T., Sassa, S., Kaiser, E.T., (1987) J. Am. Chem. Soc., 109, pp. 606-607; Radziejewski, C., Ballou, D.P., Kaiser, E.T., (1985) J. Am Chem. Soc., 107, pp. 3352-3354; Bech, L.M., Breddam, K., (1988) Carlsberg Res. Commun., 53, pp. 381-393; Bech, L.M., SÞrensen, S.B., Breddam, K., (1993) Biochemistry, 32, pp. 2845-2854; Gloss, L.M., Kirsch, J.F., (1995) Biochemistry, 34, pp. 12323-12332; Smith, H.B., Hartman, F.C., (1988) J. Biol. Chem., 263, pp. 4921-4925; Wynn, R., Harkins, P.C., Richards, F.M., Fox, R.O., (1996) Protein Sci., 5, pp. 1026-1031; Berglund, P., Stabile, M.R., Gold, M., Jones, J.B., Mitchinson, C., Bott, R.R., Graycar, T.P., (1996) Bioorg. Med. Chem. Lett., 6, pp. 2507-2512; Berglund, P., Desantis, G., Stabile, M.R., Shang, X., Gold, M., Bott, R.R., Graycar, T.P., Jones, J.B., (1997) J. Am. Chem. Soc., 119, pp. 5265-5266; Kenyon, G.L., Bruice, T.W., (1977) Methods Enzymol., 47, pp. 407-430; Wynn, R., Richards, F.M., (1995) Methods Enzymol., 251, pp. 351-356; Wong, C.-H., Whitesides, G.M., (1994) Enzymes in Synthetic Organic Chemistry, , Pergamon, Oxford, U.K; Power, S.D., Adams, R.M., Wells, J.A., (1986) Proc. Natl. Acad. Sci. U.S.A., 83, pp. 3096-3100; Von Der Osten, C., Branner, S., Hastrup, S., Hedegaard, L., Rasmussen, M.D., Bisgard-Frantzen, H., Carlsen, S., Mikkelsen, J.M., (1993) J. Biotechnol., 28 (1), pp. 55-68; Betzel, C., Klupsch, S., Papendorf, G., Hastrup, S., Branner, S., Wilson, K.S., (1992) J. Mol. Biol., 223, pp. 427-445; Stabile, M.R., Lai, G.W., DeSantis, G., Gold, M., Jones, J.B., Mitchinson, C., Bott, R.R., Liu, C.-C., (1996) Bioorg. Med. Chem. Lett., 6, pp. 2501-2506; GrÞn, H., Meldal, M., Breddam, K., (1992) Biochemistry, 31, pp. 6011-6018; Maurer, K.H., Markgraf, M., Goddette, D., (1996) Adv. Exp. Med. Biol., 379, pp. 243-256; Egmond, M.R., Antheunisse, W.P., Ravestein, P., Mooren, A.T., De Vlieg, J., (1996) Adv. Exp. Med. Biol., 379, pp. 219-228; Olsen, O.H., Pedersen, J.T., Betzel, C., Eschenburg, S., Branner, S., Hastrup, S., (1996) Adv. Exp. Med. Biol., 379, pp. 235-241; Schlechter, I., Berger, A., (1967) Biochem. Biophys. Res. Commun., 27, pp. 157-162; Bonneau, P.R., Graycar, T.P., Estell, D.A., Jones, J.B., (1991) J. Am. Chem. Soc., 113, pp. 1026-1030; Cunningham, L.W., Nuenke, B.J., (1959) J. Biol. Chem., 234, pp. 1447-1451; Ellman, G.L., Courtney, K.D., Andres, V., Featherstone, R.M., (1961) Biochem. Pharmacol., 7, pp. 88-95; Hsia, C.Y., Ganshaw, G., Paech, C., Murray, C.J., (1996) Anal. Biochem., 242, pp. 221-227; Jackson, S.E., Fersht, A.R., (1993) Biochemistry, 32, pp. 13909-13916; Russell, A.J., Fersht, A.R., (1987) Nature, 328 (6), pp. 496-500; DeSantis, G., Jones, J.B., Unpublished resultEstell, D.A., Graycar, T.P., Wells, J.A., (1985) J. Biol. Chem., 260, pp. 6518-6521; Bott, R., Ultsch, M., Wells, J., Powers, D., Burdick, D., Struble, M., Burnier, J., Power, S., (1987) ACS Symp. Ser., 334, pp. 139-147 NR 20140805

Tilgjengelig fra: 2012-02-03 Laget: 2012-02-03 Sist oppdatert: 2020-01-24bibliografisk kontrollert

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