[en] In PBP4a, a Bacillus subtilis class-C1 penicillin-binding protein (PBP), four clustered lysine
(K) residues, K86, K114, K119, and K265, protrude from domain II. Replacement of these
amino acids with glutamine (Q) residues by site-directed mutagenesis yielded Mut4KQ
PBP4a. When produced in Escherichia coli without its predicted Sec-signal peptide, wildtype
(WT) PBP4a was found mainly associated with the host cytoplasmic membrane,
whereas Mut4KQ PBP4a remained largely unbound. After purification, the capacities of the
two proteins to bind to B. subtilis membranes were compared. The results were similar to
those obtained in E. coli: in vitro, a much higher percentage of WT PBP4a than of Mut4KQ
PBP4a was found to interact with B. subtilis membranes. Immunodetection of PBP4a in B.
subtilis membrane extracts revealed that a processed form of this PBP (as indicated by its
size) associates with the B. subtilis cytoplasmic membrane. In the absence of any amphiphilic
peptide in PBP4a, the crown of positive charges on the surface of domain II is likely
responsible for the cellular localization of this PBP and its attachment to the cytoplasmic
membrane.
Sauvage, Eric ; Université de Liège > Centre d'ingénierie des protéines
Dauvin, Marjorie ; Université de Liège > Département des sciences de la vie > Physiologie et génétique bactériennes
Joris, Bernard ; Université de Liège > Département des sciences de la vie > Physiologie et génétique bactériennes
Duez, Colette ; Université de Liège > Centre d'ingénierie des protéines
Language :
English
Title :
A lysine cluster in domain II of Bacillus subtilis PBP4a plays a role in the membrane attachment of this C1-PBP
Alternative titles :
[fr] Un groupement de lysines en surface du domaine II du PBP4a de Bacillus subtilis joue un rôle dans l'attachement de ce PBP C1 à la membrane
Publication date :
13 October 2015
Journal title :
PLoS ONE
eISSN :
1932-6203
Publisher :
Public Library of Science, San Franscisco, United States - California
Peer reviewed :
Peer Reviewed verified by ORBi
Name of the research project :
Etude des PBP de classe C
Funders :
IAP P6/19 and P7/44 F.R.S.-FNRS - Fonds de la Recherche Scientifique FRIA - Fonds pour la Formation à la Recherche dans l'Industrie et dans l'Agriculture
Funding text :
This work was supported by the Belgian Program on Interuniversity Poles of Attraction by the Prime Minister’s Office, Science Policy Programming
(IAP n°P6/19 and P7/44).
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