Crystal structure of brefeldin A esterase, a bacterial homolog of the mammalian hormone-sensitive lipase.
Brefeldin A esterase (BFAE), a detoxifying enzyme isolated from Bacillus subtilis, hydrolyzes and inactivates BFA, a potent fungal inhibitor of intracellular vesicle-dependent secretory transport and poliovirus RNA replication. We have solved the crystal structure of BFAE and we discovered that the previously reported amino acid sequence was in serious error due to frame shifts in the cDNA sequence. The correct sequence, inferred from the experimentally phased electron density map, revealed that BFAE is a homolog of the mammalian hormone sensitive lipase (HSL). It is a canonical alpha/beta hydrolase with two insertions forming the substrate binding pocket. The enzyme contains a lipase-like catalytic triad, Ser 202, Asp 308 and His 338, consistent with mutational studies that implicate the homologous Ser 424, Asp 693 and His 723 in the catalytic triad in human HSL.
Wei
Y
Y
Department of Molecular Physiology and Biological Physics, University of Virginia, Charlottesville 22906-0011, USA.
Contreras
J A
JA
Sheffield
P
P
Osterlund
T
T
Derewenda
U
U
Kneusel
R E
RE
Matern
U
U
Holm
C
C
Derewenda
Z S
ZS
D000595:Amino Acid Sequence; D000818:Animals; D001412:Bacillus subtilis / Q000737:chemistry / Q000201:enzymology*; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D001483:Base Sequence; D002265:Carboxylic Ester Hydrolases / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism*; D020134:Catalytic Domain; D018360:Crystallography, X-Ray; D006728:Hormones / Q000494:pharmacology; D006801:Humans; D008322:Mammals; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011487:Protein Conformation; D017510:Protein Folding; D015203:Reproducibility of Results; D017421:Sequence Analysis / Q000379:methods; D017386:Sequence Homology, Amino Acid; D002787:Sterol Esterase / Q000187:drug effects / Q000378:metabolism
D001426:Bacterial Proteins; D006728:Hormones; D002265:Carboxylic Ester Hydrolases; C085832:brefeldin A esterase; D002787:Sterol Esterase