The molybdenum site of sulfite oxidase: A comparison of wild-type and the cysteine 207 to serine mutant using X-ray absorption spectroscopy

Journal of the American Chemical Society

Volumn 118, Issue 36, 1996, Pages 8588-8592

  George, Graham N ^a   Garrett, Robert M ^b   Prince, Roger C ^c   Rajagopalan, K V ^b  

^a SLAC NATIONAL ACCELERATOR LABORATORY   (United States)

^b DUKE UNIVERSITY SCHOOL OF MEDICINE   (United States)

^c EXXONMOBIL UPSTREAM RESEARCH COMPANY   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

SULFITE OXIDASE;

ARTICLE; ENZYME ACTIVE SITE; ENZYME ANALYSIS; HUMAN; MUTANT; X RAY ANALYSIS;

EID: 0029816512     PISSN: 00027863     EISSN: None     Source Type: Journal    
DOI: 10.1021/ja961218h     Document Type: Article

References (49)

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30. As expected, complexes possessing the Mo≡N group also show a very similar near-edge feature (Laplaza, C. E.; Johnson, M. J. A.; Peters, J.; Odom, A. L.; Kim, E.; Cummins, C. C.; George, G. N.; Pickering, I. J. Submitted for publication).
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35. We also considered the possibility that the mutant protein might be unstable, perhaps losing molybdenum as molybdate during the preparation of concentrated EXAFS samples. If this were the case then an apparent increase in Mo=O coordination number might be observed due to molybdate contamination. In an attempt to eliminate this possibility, a sample of the mutant enzyme was rapidly gel-filtered on a Sephadex G100 column, with the eluant flowing directly into an EXAFS sample cuvette, which was then frozen and the XAS measured immediately. The Mo K-edge near-edge spectrum of enzyme thus treated was found to be identical to that shown in Figure 2, and we conclude that there is no significant contamination with low molecular weight species. The Mo K-edge XAS spectrum of the mutant protein also proved to be totally reproducible from preparation to preparation.
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41. 10
42. -1, and thus, given accurate normalization, should be of equivalent reliability.
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