Extensive comparison of the substrate preferences of two subtilisins as determined with peptide substrates which are based on the principle of intramolecular quenching - PubMed (original) (raw)
Comparative Study
. 1992 Jul 7;31(26):6011-8.
doi: 10.1021/bi00141a008.
Affiliations
- PMID: 1627543
- DOI: 10.1021/bi00141a008
Comparative Study
Extensive comparison of the substrate preferences of two subtilisins as determined with peptide substrates which are based on the principle of intramolecular quenching
H Grøn et al. Biochemistry. 1992.
Abstract
Subtilisins are serine endopeptidases with an extended binding cleft comprising at least eight binding subsites. Interestingly, subsites distant from the scissile bond play a dominant role in determining the specificity of the enzymes. The development of internally quenched fluorogenic substrates, which allow polypeptides of more than 11 amino acids to be inserted between the donor and the acceptor, has rendered it possible to perform a highly systematic mapping of the individual subsites of the active sites of subtilisin BPN' from Bacillus amyloliquefaciens and Savinase from Bacillus lentus. For each enzyme, the eight positions S5-S'3 were characterized by determination of kcat/KM values for the hydrolysis of substrates in which the amino acids were systematically varied. The results emphasize that in both subtilisin BPN' and Savinase interactions between substrate and S4 and S1 are very important. However, it is apparent that interactions between other subsites and the substrate exert a significant influence on the substrate preference. The results are rationalized on the basis of the structural data available for the two enzymes.
Similar articles
- Interdependency of the binding subsites in subtilisin.
Grøn H, Breddam K. Grøn H, et al. Biochemistry. 1992 Sep 22;31(37):8967-71. doi: 10.1021/bi00152a037. Biochemistry. 1992. PMID: 1390683 - Significance of hydrophobic S4-P4 interactions in subtilisin 309 from Bacillus lentus.
Bech LM, Sørensen SB, Breddam K. Bech LM, et al. Biochemistry. 1993 Mar 23;32(11):2845-52. doi: 10.1021/bi00062a016. Biochemistry. 1993. PMID: 8457550 - Crystal structure of the alkaline proteinase Savinase from Bacillus lentus at 1.4 A resolution.
Betzel C, Klupsch S, Papendorf G, Hastrup S, Branner S, Wilson KS. Betzel C, et al. J Mol Biol. 1992 Jan 20;223(2):427-45. doi: 10.1016/0022-2836(92)90662-4. J Mol Biol. 1992. PMID: 1738156 - Kinetics of subtilisin and thiolsubtilisin.
Philipp M, Bender ML. Philipp M, et al. Mol Cell Biochem. 1983;51(1):5-32. doi: 10.1007/BF00215583. Mol Cell Biochem. 1983. PMID: 6343835 Review. - Subtilisin--an enzyme designed to be engineered.
Wells JA, Estell DA. Wells JA, et al. Trends Biochem Sci. 1988 Aug;13(8):291-7. doi: 10.1016/0968-0004(88)90121-1. Trends Biochem Sci. 1988. PMID: 3154281 Review. No abstract available.
Cited by
- In Silico Strategies to Predict Anti-aging Features of Whey Peptides.
Rama GR, Saraiva Macedo Timmers LF, Volken de Souza CF. Rama GR, et al. Mol Biotechnol. 2024 Sep;66(9):2426-2440. doi: 10.1007/s12033-023-00887-9. Epub 2023 Sep 22. Mol Biotechnol. 2024. PMID: 37737930 - New robust subtilisins from halotolerant and halophilic Bacillaceae.
Falkenberg F, Voß L, Bott M, Bongaerts J, Siegert P. Falkenberg F, et al. Appl Microbiol Biotechnol. 2023 Jun;107(12):3939-3954. doi: 10.1007/s00253-023-12553-w. Epub 2023 May 9. Appl Microbiol Biotechnol. 2023. PMID: 37160606 Free PMC article. - Characterization of a novel cold-adapted intracellular serine protease from the extremophile Planococcus halocryophilus Or1.
Rasmussen CB, Scavenius C, Thøgersen IB, Harwood SL, Larsen Ø, Bjerga GEK, Stougaard P, Enghild JJ, Thøgersen MS. Rasmussen CB, et al. Front Microbiol. 2023 Feb 23;14:1121857. doi: 10.3389/fmicb.2023.1121857. eCollection 2023. Front Microbiol. 2023. PMID: 36910232 Free PMC article. - Challenges in the use of sortase and other peptide ligases for site-specific protein modification.
Morgan HE, Turnbull WB, Webb ME. Morgan HE, et al. Chem Soc Rev. 2022 May 23;51(10):4121-4145. doi: 10.1039/d0cs01148g. Chem Soc Rev. 2022. PMID: 35510539 Free PMC article. Review. - Destruction of Staphylococcus aureus biofilms by combining an antibiotic with subtilisin A or calcium gluconate.
Liu J, Madec JY, Bousquet-Mélou A, Haenni M, Ferran AA. Liu J, et al. Sci Rep. 2021 Mar 18;11(1):6225. doi: 10.1038/s41598-021-85722-4. Sci Rep. 2021. PMID: 33737602 Free PMC article.
Publication types
MeSH terms
Substances
LinkOut - more resources
Other Literature Sources