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Journal of Bacteriology, October 1998, p. 5313-5318, Vol. 180, No. 20
0021-9193/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Conserved Structural Regions Involved in the Catalytic Mechanism of Escherichia coli K-12 WaaO (RfaI)

Keigo Shibayama,1 Shinji Ohsuka,1 Toshihiko Tanaka,1 Yoshichika Arakawa,2 and Michio Ohta1,*

Department of Bacteriology, School of Medicine, Nagoya University, Nagoya, 466-8550,1 and Department of Bacterial and Blood Products, National Institute of Infectious Diseases, Tokyo, 208-0011,2 Japan

Received 14 May 1998/Accepted 10 August 1998

Escherichia coli K-12 WaaO (formerly known as RfaI) is a nonprocessive alpha -1,3 glucosyltransferase, involved in the synthesis of the R core of lipopolysaccharide. By comparing the amino acid sequence of WaaO with those of 11 homologous alpha -glycosyltransferases, four strictly conserved regions, I, II, III, and IV, were identified. Since functionally related transferases are predicted to have a similar architecture in the catalytic sites, it is assumed that these four regions are directly involved in the formation of alpha -glycosidic linkage from alpha -linked nucleotide diphospho-sugar donor. Hydrophobic cluster analysis revealed a conserved domain at the N termini of these alpha -glycosyltransferases. This domain was similar to that previously reported for beta -glycosyltransferases. Thus, this domain is likely to be involved in the formation of beta -glycosidic linkage between the donor sugar and the enzyme at the first step of the reaction. Site-directed mutagenesis analysis of E. coli K-12 WaaO revealed four critical amino acid residues.


* Corresponding author. Mailing address: Department of Bacteriology, Nagoya University, School of Medicine, 65 Tsurumai, Showa, Nagoya, Aichi, 466-8550, Japan. Phone: 81-52-744-2103. Fax: 81-52-744-2107. E-mail: mohta{at}tsuru.med.nagoya-u.ac.jp.


Journal of Bacteriology, October 1998, p. 5313-5318, Vol. 180, No. 20
0021-9193/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.



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