ATP sulfurylases from sulfate-reducing bacteria of the genus Desulfovibrio. A novel metalloprotein containing cobalt and zinc.
Adenosine triphosphate sulfurylase catalyzes the formation of adenosine 5'-phosphosulfate from adenosine triphosphate and sulfate. The enzyme plays a crucial role in sulfate activation, the key step for sulfate utilization, and has been purified from crude extracts of Desulfovibrio desulfuricans ATCC 27774 and Desulfovibrio gigas. Both proteins are homotrimers [141 kDa (3 x 47) for D. desulfuricans and 147 kDa (3 x 49) for D. gigas] and have been identified, for the first time, as metalloproteins containing cobalt and zinc. EXAFS reveals that either cobalt or zinc binds endogenously at presumably equivalent metal binding sites and is tetrahedrally coordinated to one nitrogen and three sulfur atoms. Furthermore, the electronic absorption spectra display charge-transfer bands at 335 and 370 nm consistent with sulfur coordination to cobalt, and as expected for a distorted tetrahedral cobalt geometry, d-d bands are observed at 625, 666, and 715 nm. This geometry is supported by the observation of high-spin Co2+ EPR signals at g approximately 6.5.
Gavel
O Y
OY
Departamento de Química, Centro de Química Fina e Biotecnologia, Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, Portugal.
Bursakov
S A
SA
Calvete
J J
JJ
George
G N
GN
Moura
J J
JJ
Moura
I
I
D000595:Amino Acid Sequence; D001665:Binding Sites; D003035:Cobalt / Q000737:chemistry* / Q000378:metabolism; D003901:Desulfovibrio / Q000201:enzymology*; D004578:Electron Spin Resonance Spectroscopy; D008667:Metalloproteins / Q000737:chemistry* / Q000378:metabolism; D008969:Molecular Sequence Data; D010084:Oxidation-Reduction; D013056:Spectrophotometry, Ultraviolet; D013057:Spectrum Analysis; D013430:Sulfate Adenylyltransferase / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D013431:Sulfates / Q000737:chemistry; D014965:X-Rays; D015032:Zinc / Q000737:chemistry* / Q000378:metabolism
D008667:Metalloproteins; D013431:Sulfates; D003035:Cobalt; D013430:Sulfate Adenylyltransferase; D015032:Zinc