Structural Analysis of Escherichia coli OpgG, a Protein Required for the Biosynthesis of Osmoregulated Periplasmic Glucans

• Published in: Journal of molecular biology Vol. 342; no. 1; pp. 195 - 205
• Main Authors: Hanoulle, Xavier, Rollet, Eglantine, Clantin, Bernard, Landrieu, Isabelle, Ödberg-Ferragut, Carmen, Lippens, Guy, Bohin, Jean-Pierre, Villeret, Vincent
• Format: Journal Article
• Language: English
• Published: England Elsevier Ltd 03.09.2004; Elsevier
• Subjects: Amino Acid Sequence; Biochemistry; Biochemistry, Molecular Biology; biosynthesis; branching enzymes; carbohydrates; Crystallography, X-Ray; Escherichia coli; Escherichia coli - metabolism; Escherichia coli Proteins - chemistry; Escherichia coli Proteins - genetics; Glucans - biosynthesis; Glucans - chemistry; GTP-Binding Proteins - chemistry; GTP-Binding Proteins - genetics; Life Sciences; Microbiology and Parasitology; Models, Molecular; Molecular Sequence Data; periplasmic osmoregulated glucans; Periplasmic Proteins - biosynthesis; Periplasmic Proteins - chemistry; Protein Structure, Secondary; Protein Structure, Tertiary; Proteobacteria; Selenomethionine - chemistry; X-ray structure; carbohydrates; mBE; MAD; periplasmic osmoregulated glucans; branching enzymes; SeMet-OpgG; biosynthesis; OPGs; EPS; X-ray structure; Ig
• ISSN: 0022-2836; 1089-8638
• DOI: 10.1016/j.jmb.2004.07.004

Osmoregulated periplasmic glucans (OPGs) G protein (OpgG) is required for OPGs biosynthesis. OPGs from Escherichia coli are branched glucans, with a backbone of β-1,2 glucose units and with branches attached by β-1,6 linkages. In Proteobacteria, OPGs are involved in osmoprotection, biofilm formation, virulence and resistance to antibiotics. Despite their important biological implications, enzymes synthesizing OPGs are poorly characterized. Here, we report the 2.5 Å crystal structure of OpgG from E. coli . The structure was solved using a selenemethionine derivative of OpgG and the multiple anomalous diffraction method (MAD). The protein is composed of two β-sandwich domains connected by one turn of 3 10 helix. The N-terminal domain (residues 22–388) displays a 25-stranded β-sandwich fold found in several carbohydrate-related proteins. It exhibits a large cleft comprising many aromatic and acidic residues. This putative binding site shares some similarities with enzymes such as galactose mutarotase and glucodextranase, suggesting a potential catalytic role for this domain in OPG synthesis. On the other hand, the C-terminal domain (residues 401–512) has a seven-stranded immunoglobulin-like β-sandwich fold, found in many proteins where it is mainly implicated in interactions with other molecules. The structural data suggest that OpgG is an OPG branching enzyme in which the catalytic activity is located in the large N-terminal domain and controlled via the smaller C-terminal domain.