Selective 15N labeling and direct observation by NMR of the active-site glutamine of Fe-containing superoxide dismutase

Carrie K. Vance; Young M. Kang; Anne Frances Miller

doi:10.1023/A:1018662421878

Selective ¹⁵N labeling and direct observation by NMR of the active-site glutamine of Fe-containing superoxide dismutase

Carrie K. Vance, Young M. Kang, Anne Frances Miller

Research output: Contribution to journal › Article › peer-review

13 Scopus citations

Abstract

The glutamine in position 69 is one of only three conserved active-site amino acid differences between Fe- and Mn-containing superoxide dismutases (SODs). We have refined the conditions for extremely selective labeling of the side chains of glutamine with ¹⁵N, and thus obtained dramatically simplified spectra, despite the large size of Fe-SOD. The improved resolution afforded by such highly specific labeling permits the use of direct ¹⁵N detection to observe and assign Gln 69, even though its distance to the paramagnetic Fe²⁺ is only 5 Å. Selective glutamine side-chain labeling is inexpensive and has general utility for large (and paramagnet-containing) proteins.

Original language	English
Pages (from-to)	201-206
Number of pages	6
Journal	Journal of Biomolecular NMR
Volume	9
Issue number	2
DOIs	https://doi.org/10.1023/A:1018662421878
State	Published - 1997

Bibliographical note

Funding Information:
This work was supported by N.S.F. grant MCB-9418181, the Petroleum Research Foundation ACS-PRF 28379-G4, and an institutional research grant from the American Cancer Society, IRG 11-33. C.K.V. is supported by NIGMS/N.I.H. training grant T32-GM07231.

Keywords

Glutamine
N
Selective labeling
Superoxide dismutase

ASJC Scopus subject areas

Biochemistry
Spectroscopy

Access to Document

10.1023/A:1018662421878

Cite this

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title = "Selective 15N labeling and direct observation by NMR of the active-site glutamine of Fe-containing superoxide dismutase",

abstract = "The glutamine in position 69 is one of only three conserved active-site amino acid differences between Fe- and Mn-containing superoxide dismutases (SODs). We have refined the conditions for extremely selective labeling of the side chains of glutamine with 15N, and thus obtained dramatically simplified spectra, despite the large size of Fe-SOD. The improved resolution afforded by such highly specific labeling permits the use of direct 15N detection to observe and assign Gln 69, even though its distance to the paramagnetic Fe2+ is only 5 {\AA}. Selective glutamine side-chain labeling is inexpensive and has general utility for large (and paramagnet-containing) proteins.",

keywords = "Glutamine, N, Selective labeling, Superoxide dismutase",

author = "Vance, {Carrie K.} and Kang, {Young M.} and Miller, {Anne Frances}",

note = "Funding Information: This work was supported by N.S.F. grant MCB-9418181, the Petroleum Research Foundation ACS-PRF 28379-G4, and an institutional research grant from the American Cancer Society, IRG 11-33. C.K.V. is supported by NIGMS/N.I.H. training grant T32-GM07231.",

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T1 - Selective 15N labeling and direct observation by NMR of the active-site glutamine of Fe-containing superoxide dismutase

AU - Vance, Carrie K.

AU - Kang, Young M.

AU - Miller, Anne Frances

N1 - Funding Information: This work was supported by N.S.F. grant MCB-9418181, the Petroleum Research Foundation ACS-PRF 28379-G4, and an institutional research grant from the American Cancer Society, IRG 11-33. C.K.V. is supported by NIGMS/N.I.H. training grant T32-GM07231.

PY - 1997

Y1 - 1997

N2 - The glutamine in position 69 is one of only three conserved active-site amino acid differences between Fe- and Mn-containing superoxide dismutases (SODs). We have refined the conditions for extremely selective labeling of the side chains of glutamine with 15N, and thus obtained dramatically simplified spectra, despite the large size of Fe-SOD. The improved resolution afforded by such highly specific labeling permits the use of direct 15N detection to observe and assign Gln 69, even though its distance to the paramagnetic Fe2+ is only 5 Å. Selective glutamine side-chain labeling is inexpensive and has general utility for large (and paramagnet-containing) proteins.

AB - The glutamine in position 69 is one of only three conserved active-site amino acid differences between Fe- and Mn-containing superoxide dismutases (SODs). We have refined the conditions for extremely selective labeling of the side chains of glutamine with 15N, and thus obtained dramatically simplified spectra, despite the large size of Fe-SOD. The improved resolution afforded by such highly specific labeling permits the use of direct 15N detection to observe and assign Gln 69, even though its distance to the paramagnetic Fe2+ is only 5 Å. Selective glutamine side-chain labeling is inexpensive and has general utility for large (and paramagnet-containing) proteins.

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KW - Selective labeling

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