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J Biol Chem, Vol. 273, Issue 29, 18586-18593, July 17, 1998
Characterization of the
N-Acetylglucosaminyltransferase Activity Involved in the
Biosynthesis of the Staphylococcus epidermidis
Polysaccharide Intercellular Adhesin
Christiane
Gerke ,
Angelika
Kraft§,
Roderich
Süßmuth¶,
Oliver
Schweitzer , and
Friedrich
Götz
From the Mikrobielle Genetik, Universität
Tübingen, Waldhäuser Strasse 70/8, the
§ Max-Planck-Institut für Entwicklungsbiologie,
Abteilung Biochemie, Spemannstrasse 35, and the ¶ Institut
für Organische Chemie, Universität Tübingen, Auf der
Morgenstelle 18, 72076 Tübingen, Germany
The polysaccharide intercellular adhesin (PIA) is
an important factor in the colonization of medical devices by
Staphylococcus epidermidis. The genes encoding PIA
production are organized in the icaADBC
(intercellular adhesion) operon. To
study the function of the individual genes, we have established an
in vitro assay with UDP-N-acetylglucosamine,
the substrate for PIA biosynthesis, and analyzed the products by
thin-layer chromatography and mass spectrometry. IcaA alone exhibited a
low N-acetylglucosaminyltransferase activity and represents
the catalytic enzyme. Coexpression of icaA with
icaD led to a significant increase in activity. The newly
identified icaD gene is located between icaA
and icaB and overlaps both genes.
N-Acetylglucosamine oligomers produced by IcaAD reached a
maximal length of 20 residues. Only when icaA and
icaD were expressed together with icaC were
oligomer chains that react with PIA-specific antiserum synthesized.
IcaA and IcaD are located in the cytoplasmic membrane, and IcaC also
has all the structural features of an integral membrane protein. These results indicate a close interaction between IcaA, IcaD, and IcaC. Tunicamycin and bacitracin did not affect the in vitro
synthesis of PIA intermediates or the complete PIA biosynthesis
in vivo, suggesting that a undecaprenyl phosphate carrier
is not involved. IcaAD represents a novel protein combination among
-glycosyltransferases.
Copyright © 1998 by The American Society for Biochemistry and Molecular Biology, Inc.

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Copyright © 1998 by the American Society for Biochemistry and Molecular Biology.
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