Biochemical Characterisation of Phage Pseudomurein Endoisopeptidases PeiW and PeiP Using Synthetic Peptides
Joint Authors
Schofield, Linley R.
Beattie, Amy K.
Tootill, Catherine M.
Dey, Debjit
Ronimus, Ron S.
Source
Issue
Vol. 2015, Issue 2015 (31 Dec. 2015), pp.1-12, 12 p.
Publisher
Hindawi Publishing Corporation
Publication Date
2015-09-21
Country of Publication
Egypt
No. of Pages
12
Main Subjects
Abstract EN
Pseudomurein endoisopeptidases cause lysis of the cell walls of methanogens by cleaving the isopeptide bond Ala-ε-Lys in the peptide chain of pseudomurein.
PeiW and PeiP are two thermostable pseudomurein endoisopeptidases encoded by phage ΨM100 of Methanothermobacter wolfei and phages ΨM1 and ΨM2 of Methanothermobacter marburgensis, respectively.
A continuous assay using synthetic peptide substrates was developed and used in the biochemical characterisation of recombinant PeiW and PeiP.
The advantages of these synthetic peptide substrates over natural substrates are sensitivity, high purity, and characterisation and the fact that they are more easily obtained than natural substrates.
In the presence of a reducing agent, purified PeiW and PeiP each showed similar activity under aerobic and anaerobic conditions.
Both enzymes required a divalent metal for activity and showed greater thermostability in the presence of Ca2+.
PeiW and PeiP involve a cysteine residue in catalysis and have a monomeric native conformation.
The kinetic parameters, KM and kcat, were determined, and the ε-isopeptide bond between alanine and lysine was confirmed as the bond lysed by these enzymes in pseudomurein.
The new assay may have wider applications for the general study of peptidases and the identification of specific methanogens susceptible to lysis by specific pseudomurein endoisopeptidases.
American Psychological Association (APA)
Schofield, Linley R.& Beattie, Amy K.& Tootill, Catherine M.& Dey, Debjit& Ronimus, Ron S.. 2015. Biochemical Characterisation of Phage Pseudomurein Endoisopeptidases PeiW and PeiP Using Synthetic Peptides. Archaea،Vol. 2015, no. 2015, pp.1-12.
https://search.emarefa.net/detail/BIM-1053932
Modern Language Association (MLA)
Schofield, Linley R.…[et al.]. Biochemical Characterisation of Phage Pseudomurein Endoisopeptidases PeiW and PeiP Using Synthetic Peptides. Archaea No. 2015 (2015), pp.1-12.
https://search.emarefa.net/detail/BIM-1053932
American Medical Association (AMA)
Schofield, Linley R.& Beattie, Amy K.& Tootill, Catherine M.& Dey, Debjit& Ronimus, Ron S.. Biochemical Characterisation of Phage Pseudomurein Endoisopeptidases PeiW and PeiP Using Synthetic Peptides. Archaea. 2015. Vol. 2015, no. 2015, pp.1-12.
https://search.emarefa.net/detail/BIM-1053932
Data Type
Journal Articles
Language
English
Notes
Includes bibliographical references
Record ID
BIM-1053932