Substrate preferences of glutamic-acid-specific endopeptidases assessed by synthetic peptide substrates based on intramolecular fluorescence quenching - PubMed (original) (raw)
Comparative Study
Substrate preferences of glutamic-acid-specific endopeptidases assessed by synthetic peptide substrates based on intramolecular fluorescence quenching
K Breddam et al. Eur J Biochem. 1992.
Free article
Abstract
The substrate preferences of the easily available Glu/Asp-specific enzymes from Staphyllococcus aureus (V8), Bacillus licheniformis and Streptomyces griseus have been extensively investigated using a series of synthetic peptide substrates, containing an N-terminal anthraniloyl group and a 3-nitrotyrosine close to the C-terminus, allowing the fluorimetric monitoring of substrate hydrolysis by the decrease in intramolecular quenching. All three enzymes hydrolysed Glu-Xaa peptide bonds approximately 1000-fold faster than Asp-Xaa bonds and they are consequently more appropriately termed Glu-specific enzymes. The difference in kcat/Km for the hydrolysis of substrates with Glu and Asp is primarily due to a difference in kcat. The enzymes appear to hydrolyse all types of Glu-Xaa bonds, although those with Xaa as Asp and, in particular, Xaa as Pro, are hydrolysed with very low rates. The influence of the nature of the amino acid residues at the substrate positions P2, P3, P4, P'1 and P'2 has been determined and it is shown that the enzyme from S. griseus exhibits the most narrow substrate preference. The results are useful in connection with fragmentation of proteins for sequencing purposes as well as for cleavage of fusion proteins.
Similar articles
- Isolation and amino acid sequence of a glutamic acid specific endopeptidase from Bacillus licheniformis.
Svendsen I, Breddam K. Svendsen I, et al. Eur J Biochem. 1992 Feb 15;204(1):165-71. doi: 10.1111/j.1432-1033.1992.tb16619.x. Eur J Biochem. 1992. PMID: 1346764 - The specificity of carboxypeptidase Y may be altered by changing the hydrophobicity of the S'1 binding pocket.
Sørensen SB, Breddam K. Sørensen SB, et al. Protein Sci. 1997 Oct;6(10):2227-32. doi: 10.1002/pro.5560061017. Protein Sci. 1997. PMID: 9336845 Free PMC article. - Purification, characterization, cloning, and expression of a glutamic acid-specific protease from Bacillus licheniformis ATCC 14580.
Kakudo S, Kikuchi N, Kitadokoro K, Fujiwara T, Nakamura E, Okamoto H, Shin M, Tamaki M, Teraoka H, Tsuzuki H, et al. Kakudo S, et al. J Biol Chem. 1992 Nov 25;267(33):23782-8. J Biol Chem. 1992. PMID: 1429718 - A rapid method for determination of endoproteinase substrate specificity: specificity of the 3C proteinase from hepatitis A virus.
Petithory JR, Masiarz FR, Kirsch JF, Santi DV, Malcolm BA. Petithory JR, et al. Proc Natl Acad Sci U S A. 1991 Dec 15;88(24):11510-4. doi: 10.1073/pnas.88.24.11510. Proc Natl Acad Sci U S A. 1991. PMID: 1662396 Free PMC article. - The specificity of clostripain from Clostridium histolyticum. Mapping the S' subsites via acyl transfer to amino acid amides and peptides.
Ullmann D, Jakubke HD. Ullmann D, et al. Eur J Biochem. 1994 Aug 1;223(3):865-72. doi: 10.1111/j.1432-1033.1994.tb19063.x. Eur J Biochem. 1994. PMID: 8055964
Cited by
- Mammalian Esterase Activity: Implications for Peptide Prodrugs.
Petri YD, Verresen R, Gutierrez CS, Kojasoy V, Zhang E, Abularrage NS, Wralstad EC, Weiser KR, Raines RT. Petri YD, et al. Biochemistry. 2024 Oct 15;63(20):2580-2593. doi: 10.1021/acs.biochem.4c00446. Epub 2024 Oct 3. Biochemistry. 2024. PMID: 39359146 Free PMC article. - Lipid environment modulates processivity and kinetics of a presenilin homolog acting on multiple substrates in vitro.
Wu Y, Thomas GM, Thomsen M, Bahri S, Lieberman RL. Wu Y, et al. J Biol Chem. 2023 Dec;299(12):105401. doi: 10.1016/j.jbc.2023.105401. Epub 2023 Oct 29. J Biol Chem. 2023. PMID: 38270390 Free PMC article. - A nemaline myopathy-linked mutation inhibits the actin-regulatory functions of tropomodulin and leiomodin.
Schultz LE, Colpan M, Smith GE Jr, Mayfield RM, Larrinaga TM, Kostyukova AS, Gregorio CC. Schultz LE, et al. Proc Natl Acad Sci U S A. 2023 Nov 21;120(47):e2315820120. doi: 10.1073/pnas.2315820120. Epub 2023 Nov 13. Proc Natl Acad Sci U S A. 2023. PMID: 37956287 Free PMC article. - Glycoengineering of EphA4 Fc leads to a unique, long-acting and broad spectrum, Eph receptor therapeutic antagonist.
Pegg CL, Cooper LT, Zhao J, Gerometta M, Smith FM, Yeh M, Bartlett PF, Gorman JJ, Boyd AW. Pegg CL, et al. Sci Rep. 2017 Jul 26;7(1):6519. doi: 10.1038/s41598-017-06685-z. Sci Rep. 2017. PMID: 28747680 Free PMC article. - Glutamyl Endopeptidases: The Puzzle of Substrate Specificity.
Demidyuk IV, Chukhontseva KN, Kostrov SV. Demidyuk IV, et al. Acta Naturae. 2017 Apr-Jun;9(2):17-33. Acta Naturae. 2017. PMID: 28740724 Free PMC article.
Publication types
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
Other Literature Sources