AcmD, a homolog of the major autolysin AcmA of Lactococcus lactis, binds to the cell wall and contributes to cell separation and autolysis.
Lactococcus lactis expresses the homologous glucosaminidases AcmB, AcmC, AcmA and AcmD. The latter two have three C-terminal LysM repeats for peptidoglycan binding. AcmD has much shorter intervening sequences separating the LysM repeats and a lower iso-electric point (4.3) than AcmA (10.3). Under standard laboratory conditions AcmD was mainly secreted into the culture supernatant. An L. lactis acmAacmD double mutant formed longer chains than the acmA single mutant, indicating that AcmD contributes to cell separation. This phenotype could be complemented by plasmid-encoded expression of AcmD in the double mutant. No clear difference in cellular lysis and protein secretion was observed between both mutants. Nevertheless, overexpression of AcmD resulted in increased autolysis when AcmA was present (as in the wild type strain) or when AcmA was added to the culture medium of an AcmA-minus strain. Possibly, AcmD is mainly active within the cell wall, at places where proper conditions are present for its binding and catalytic activity. Various fusion proteins carrying either the three LysM repeats of AcmA or AcmD were used to study and compare their cell wall binding characteristics. Whereas binding of the LysM domain of AcmA took place at pHs ranging from 4 to 8, LysM domain of AcmD seems to bind strongest at pH 4.
Department of Molecular Genetics, Groningen Biomolecular Sciences and Biotechnology Institute, University of Groningen, Groningen, The Netherlands.
Ganesh Ram R
GR
Visweswaran
Anton
A
Steen
Kees
K
Leenhouts
Monika
M
Szeliga
Beata
B
Ruban
Anne
A
Hesseling-Meinders
Bauke W
BW
Dijkstra
Oscar P
OP
Kuipers
Jan
J
Kok
Girbe
G
Buist
D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D001433:Bacteriolysis; D002473:Cell Wall / Q000378:metabolism; D006026:Glycoside Hydrolases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D006596:Hexosaminidases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D013294:Lactococcus lactis / Q000737:chemistry / Q000166:cytology* / Q000235:genetics / Q000378:metabolism*; D009154:Mutation; D010457:Peptidoglycan / Q000378:metabolism; D011485:Protein Binding; D017434:Protein Structure, Tertiary
D001426:Bacterial Proteins; D010457:Peptidoglycan; D006026:Glycoside Hydrolases; D006596:Hexosaminidases; C026255:endo-N-acetylmuramidase