JBC, Vol. 252, Issue 12, 4118-4124, Jun, 1977
Incorporation of N-acetyl-D-glucosamine from UDP-N-acetyl-D-glucosamine by isolated membranes of Bacillus subtilis. Identification of undecaprenyl poly(N-acetylglucosaminyl pyrophosphate)
G. E. Bettinger, A. N. Chatterjee and F. E. Young
Membrane isolated from Bacillus subtilis strain 168 incorporated GlcNAc
from UDP-GlcNAc directly onto undecaprenyl phosphate via
transphosphorylation and subsequent transglucosylations. Chain lengths of
6, 4, and 1 units of GlcNAc were found. Approximately 80% of the isotope
incorporated was extracted into chloroform:methanol (2:1 v/v), and could be
distinguished from the undecaprenyl disaccharide cell wall intermediate by
a different elution pattern on DEAE-cellulose (acetate form). The
GlcNAc-lipid(s) were eluted from a similar column in
chloroform:methanol:water (10:10:3, v/v) with 6 mM NH4COOH indicating a
pyrophosphate linkage between the lipid and the GlcNAc. The GlcNAc-lipid(s)
were not degraded by conditions which completely deacylated [32P]glyceryl
phospholipids, but were rapidly hydrolyzed by mild acid treatment (0.005 N
HCl, 90 degrees) with the release of oligosaccharide phosphate (typical of
sugars linked to undecaprenyl pyrophosphate). Catalytic hydrogenation of
the GlcNAc-lipid(s) resulted in the release of water-soluble sugar
phosphate. Under these same conditions, undecaprenyl pyrophosphate and
undecaprenyl disaccharide cell wall intermediate were similarly effected
while [32P]glyceryl phospholipids remained intact. The formation of
GlcNAc-lipid(s) in vitro was inhibited if membranes were prepared from
cells previously treated with bacitracin. Thus, the GlcNAc-lipid(s) has the
properties of undecaprenyl poly(N-acetylglucosaminyl pyrophosphate) and may
represent a new synthetic role of the polyisoprenyl lipid in B. subtilis.
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