Introduction 1 2 Bordetella parapertussis Bordetella bronchiseptica B. bronchiseptica 3 4 B. parapertussis 5 B. parapertussis 6 B. bronchiseptica B. parapertussis 7 B. bronchiseptica wlb bpl 8 B. bronchiseptica B. parapertussis l l 9 in vitro 10 wbm wlb wbm wbmF wbmG wbmH wbm wbm wbmF wbmG wbmH wbm l d d l galacto d d 11 12 13–16 d l l d wbmF wbmG wbmH B. bronchiseptica 17 wbm in vivo wbm Results wbmF wbmG wbmH wbmF wbmG wbmH Fig. 1 wbmG wbmH wbmF in trans Fig. 1 flaA wbmG wbmH wbmF wbmF wbmF wbm wbmF 18 The gene products WbmF, WbmG and WbmH are all in the extended SDR family wbmF wbmG wbmH 2 Fig. 2 Escherichia coli Streptomyces venezuelae Table 1 19 Fig. 2 6 6 6 wbm 20 Table 2 6 6 6 Fig. 3 Fig. 3 6 6 2 2 2 Cofactor binding 6 6 E. coli 6 20 6 21 22 via 6 Binding of substrate 6 6 Fig. 3 6 Fig. 3 Fig. 3 + 6 Fig. 4 S. venezuelae Fig. 4 23 1R6D Fig. 3 Fig. 3 E. coli 24 S. venezuelae 23 Pseudomonas aeruginosa 25 n 2 Fig. 5 Potential catalytic chemistry 23 26 Fig. 6 Bordetella wbm 21 12 Fig. 6 cis Modelling of the putative substrate into the WbmF active site d l d d Fig. 7 Discussion wbmF wbmG wbmH d l wbm 27 E. coli 1BWS 28 20 29 d manno l galacto + 12 30 K a 23 E. coli k cat 31 E. coli k cat 32 d Fig. 7 Fig. 8 d K a Aspergillus nidulans 33 34 P. aeruginosa 26 34 Fig. 8 35 wbmF 36 12 l wbmF l Streptomyces spheroides l 37 d xylo in vitro k cat 38 39 in vitro d in vitro Materials and Methods Bacterial strains, plasmids and culture conditions Table 3 Bordetella E. coli − 1 − 1 − 1 E. coli − 1 B. bronchiseptica − 1 40 r 41 B. bronchiseptica 4 DNA methods 42 Taq wbmF wbmG wbmH wbmF wbmF B. bronchiseptica 43 Tc wbmG TCTAGA wbmG wbmG Tc wbmH in vitro wbm 18 AJ007747 wbmH E. coli B. bronchiseptica E. coli 44 sacB B. bronchiseptica 45 Complementation of mutations B. bronchiseptica flaA TCTAGA GGATCCCATATG wbmF CATATG AAGCTT wbmF wbmG CATATG AAGCTT wbmH CATATG AAGCTT B. bronchiseptica E. coli 46 trans E. coli 44,47 SDS-PAGE, silver stain and immunoblot analysis of LPS B. bronchiseptica 48 49 50 51 B. bronchiseptica 35 X-ray crystallography 20 52 53 54 55 20 56 57 58 59 60 61 62 63 64 65 Table 2 66 57 67 68 Supplementary Data Modelling of putative substrate into WbmF structure d 58 57 d d 69 Protein Data Bank accession codes 2PZJ 2Q1T 2Q1U 2Q1S 2PZK 2PZL 2PZM 2Q1W Supplementary data Supplementary Data  Supplementary Data doi:10.1016/j.jmb.2007.09.055