In vitro synthesis of peptidoglycan by spheroplasts of Proteus mirabilis grown in the presence of penicillin.
Spheroplasts of the unstable L-form of Proteus mirabilis with fragile, shape defective cell walls grown in medium containing 120 mg/l penicillin G and then killed and permeabilized by ether treatment, were capable of in vitro synthesis of peptidoglycan from the precursors UDP-GlcNAc and UDP-MurNAc-L-Ala-D-Glu(ms-A2pm-D-Ala-D-Ala). The in vitro peptidoglycan was extensively peptide-crosslinked, indicating a continuing function of peptidoglycan transpeptidase in the spheroplasts. The seven penicillin-binding proteins (PBPs) of P. mirabilis with their functions as multiple peptidoglycan transpeptidases were shown to be saturated in the spheroplasts and thereby functionally inactivated by the penicillin of the growth medium to a very different degree. Complete or almost complete saturation occurred with the PBPs 1A, 1B, and 3, for which functions as indispensable transpeptidases in Escherichia coli have been postulated. In contrast, PBPs 5 and 6 were not saturated in the L-form spheroplasts. Transpeptidase function has been described previously in PBP 5 of P. mirabilis. The working hypothesis is proposed that synthesis of the functionally defective peptidoglycan of L-form spheroplasts in the presence of penicillin takes place with transpeptidase function of PBP 5.
D000107:Acetylation; D001426:Bacterial Proteins; D002352:Carrier Proteins / Q000032:analysis; D006602:Hexosyltransferases; D066298:In Vitro Techniques; D002267:Muramoylpentapeptide Carboxypeptidase / Q000032:analysis; D046915:Penicillin-Binding Proteins; D010406:Penicillins / Q000494:pharmacology*; D010457:Peptidoglycan / Q000096:biosynthesis*; D010458:Peptidyl Transferases / Q000378:metabolism; D011513:Proteus mirabilis / Q000378:metabolism*; D013104:Spheroplasts / Q000378:metabolism
D001426:Bacterial Proteins; D002352:Carrier Proteins; D046915:Penicillin-Binding Proteins; D010406:Penicillins; D010457:Peptidoglycan; D010458:Peptidyl Transferases; D006602:Hexosyltransferases; D002267:Muramoylpentapeptide Carboxypeptidase