D-alanylation of lipoteichoic acid: role of the D-alanyl carrier protein in acylation.
The D-alanylation of membrane-associated lipoteichoic acid (LTA) in gram-positive organisms requires the D-alanine-D-alanyl carrier protein ligase (AMP) (Dcl) and the D-alanyl carrier protein (Dcp). The dlt operon encoding these proteins (dltA and dltC) also includes dltB and dltD. dltB encodes a putative transport system, while dltD encodes a protein which facilitates the binding of Dcp and Dcl for ligation with D-alanine and has thioesterase activity for mischarged D-alanyl-acyl carrier proteins (ACPs). In previous results it was shown that D-alanyl-Dcp donates its ester residue to membrane-associated LTA (M. P. Heaton and F. C. Neuhaus, J. Bacteriol. 176: 681-690, 1994). However, all efforts to identify an enzyme which catalyzes this D-alanylation process were unsuccessful. It was discovered that incubation of D-alanyl-Dcp in the presence of LTA resulted in the time-dependent hydrolysis of this D-alanyl thioester. D-Alanyl-ACP in the presence of LTA was not hydrolyzed. When Dcp was incubated with membrane-associated D-alanyl LTA, a time and concentration-dependent formation of D-alanyl-Dcp was found. The addition of NaCl to this reaction inhibited the formation of D-alanyl-Dcp and stimulated the hydrolysis of D-alanyl-Dcp. Since these reactions are specific for the carrier protein (Dcp), it is suggested that Dcp has a unique binding site which interacts with the poly(Gro-P) moiety of LTA. It is this specific interaction that provides the functional specificity for the D-alanylation process. The reversibility of this process provides a mechanism for the transacylation of the D-alanyl ester residues between LTA and wall teichoic acid.
Department of Biochemistry, Molecular Biology, and Cell Biology, Northwestern University, Evanston, Illinois 60208-3500, USA.
M Y
MY
Kiriukhin
F C
FC
Neuhaus
D000409:Alanine / Q000378:metabolism*; D001426:Bacterial Proteins; D002250:Carbon Radioisotopes / Q000378:metabolism; D002352:Carrier Proteins / Q000378:metabolism*; D002462:Cell Membrane / Q000378:metabolism; D006868:Hydrolysis; D007780:Lacticaseibacillus casei / Q000378:metabolism*; D008070:Lipopolysaccharides / Q000378:metabolism*; D012965:Sodium Chloride / Q000494:pharmacology; D013682:Teichoic Acids / Q000378:metabolism*
D001426:Bacterial Proteins; D002250:Carbon Radioisotopes; D002352:Carrier Proteins; C085651:D-alanyl carrier protein, bacteria; D008070:Lipopolysaccharides; D013682:Teichoic Acids; D012965:Sodium Chloride; C009900:lipoteichoic acid; D000409:Alanine