Ethanolamine ammonia-lyase has a 'base-on' binding mode for coenzyme B12

Andreas Abend; Vahe Bandarian; Rainer Nitsche; Erhard Stupperich; Janos Rétey; George H. Reed

doi:10.1006/abbi.1999.1382

Ethanolamine ammonia-lyase has a 'base-on' binding mode for coenzyme B₁₂

Andreas Abend, Vahe Bandarian, Rainer Nitsche, Erhard Stupperich, Janos Rétey, George H. Reed

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

65 Scopus citations

Abstract

Ethanolamine ammonia-lyase (EAL, EC 4.3.1.7) catalyzes a coenzyme B₁₂- dependent deamination of vicinal amino alcohols. The mode of binding of coenzyme B₁₂ to EAL has been investigated by electron paramagnetic resonance spectroscopy (EPR) using [¹⁵N]-dimethylbenzimidazole-coenzyme B₁₂. EAL was incubated with either unlabeled or ¹⁵N-enriched coenzyme B₁₂ and then either exposed to light or treated with ethanol to generate the cleaved form of the cofactor, cob(II)alamin (B₁₂(r)) bound in the active site. The reaction mixtures were examined by EPR spectroscopy at 77 K. ¹⁵N superhyperfine splitting in the EPR signals of the low-spin Co²⁺ of B₁₂(r), bound in the active site of EAL, indicates that the dimethylbenzimidazole moiety of the cofactor contributes the lower axial ligand consistent with 'base-on' binding of coenzyme B₁₂ to EAL.

Original language	English (US)
Pages (from-to)	138-141
Number of pages	4
Journal	Archives of Biochemistry and Biophysics
Volume	370
Issue number	1
DOIs	https://doi.org/10.1006/abbi.1999.1382
State	Published - Oct 1 1999
Externally published	Yes

ASJC Scopus subject areas

Biophysics
Biochemistry
Molecular Biology

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10.1006/abbi.1999.1382

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@article{1cd949d5ea114b22959f8e7d916a2ecd,

title = "Ethanolamine ammonia-lyase has a 'base-on' binding mode for coenzyme B12",

abstract = "Ethanolamine ammonia-lyase (EAL, EC 4.3.1.7) catalyzes a coenzyme B12- dependent deamination of vicinal amino alcohols. The mode of binding of coenzyme B12 to EAL has been investigated by electron paramagnetic resonance spectroscopy (EPR) using [15N]-dimethylbenzimidazole-coenzyme B12. EAL was incubated with either unlabeled or 15N-enriched coenzyme B12 and then either exposed to light or treated with ethanol to generate the cleaved form of the cofactor, cob(II)alamin (B12(r)) bound in the active site. The reaction mixtures were examined by EPR spectroscopy at 77 K. 15N superhyperfine splitting in the EPR signals of the low-spin Co2+ of B12(r), bound in the active site of EAL, indicates that the dimethylbenzimidazole moiety of the cofactor contributes the lower axial ligand consistent with 'base-on' binding of coenzyme B12 to EAL.",

author = "Andreas Abend and Vahe Bandarian and Rainer Nitsche and Erhard Stupperich and Janos R{\'e}tey and Reed, {George H.}",

note = "Funding Information: 1This work was supported in part by NIH Grant GM35752 (G.H.R.), Deutsche Forschungsgemeinschaft (J.R. and postdoctoral fellowship to A.A.), and the European Communion (R.N. and J.R.). 2Present address: Merck & Company, Inc., Sumneytown Pike, P.O. Box 4, West Point, PA 19486-0004. 3Present address: University of Michigan, Biophys. Res. Div., Chem. Sci. Bldg., 930 N. University Ave., Ann Arbor, MI 48109-1055. 4To whom correspondence should be addressed. Institute for Enzyme Research, University of Wisconsin—Madison, 1710 University Ave., Madison, WI 53705. Fax: 608-265-2904. E-mail: reed@enzyme. wisc.edu. 5Abbreviations used: EAL, ethanolamine ammonia-lyase; EPR, electron paramagnetic resonance; coenzyme B12, adenosylcobalamin; B12r, cob(II)alamin; Hepes, N-2-hydroxyethylpiperazine-N′-2-ethanesulfonic acid.",

year = "1999",

month = oct,

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doi = "10.1006/abbi.1999.1382",

language = "English (US)",

volume = "370",

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journal = "Archives of Biochemistry and Biophysics",

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T1 - Ethanolamine ammonia-lyase has a 'base-on' binding mode for coenzyme B12

AU - Abend, Andreas

AU - Bandarian, Vahe

AU - Nitsche, Rainer

AU - Stupperich, Erhard

AU - Rétey, Janos

AU - Reed, George H.

N1 - Funding Information: 1This work was supported in part by NIH Grant GM35752 (G.H.R.), Deutsche Forschungsgemeinschaft (J.R. and postdoctoral fellowship to A.A.), and the European Communion (R.N. and J.R.). 2Present address: Merck & Company, Inc., Sumneytown Pike, P.O. Box 4, West Point, PA 19486-0004. 3Present address: University of Michigan, Biophys. Res. Div., Chem. Sci. Bldg., 930 N. University Ave., Ann Arbor, MI 48109-1055. 4To whom correspondence should be addressed. Institute for Enzyme Research, University of Wisconsin—Madison, 1710 University Ave., Madison, WI 53705. Fax: 608-265-2904. E-mail: reed@enzyme. wisc.edu. 5Abbreviations used: EAL, ethanolamine ammonia-lyase; EPR, electron paramagnetic resonance; coenzyme B12, adenosylcobalamin; B12r, cob(II)alamin; Hepes, N-2-hydroxyethylpiperazine-N′-2-ethanesulfonic acid.

PY - 1999/10/1

Y1 - 1999/10/1

N2 - Ethanolamine ammonia-lyase (EAL, EC 4.3.1.7) catalyzes a coenzyme B12- dependent deamination of vicinal amino alcohols. The mode of binding of coenzyme B12 to EAL has been investigated by electron paramagnetic resonance spectroscopy (EPR) using [15N]-dimethylbenzimidazole-coenzyme B12. EAL was incubated with either unlabeled or 15N-enriched coenzyme B12 and then either exposed to light or treated with ethanol to generate the cleaved form of the cofactor, cob(II)alamin (B12(r)) bound in the active site. The reaction mixtures were examined by EPR spectroscopy at 77 K. 15N superhyperfine splitting in the EPR signals of the low-spin Co2+ of B12(r), bound in the active site of EAL, indicates that the dimethylbenzimidazole moiety of the cofactor contributes the lower axial ligand consistent with 'base-on' binding of coenzyme B12 to EAL.

AB - Ethanolamine ammonia-lyase (EAL, EC 4.3.1.7) catalyzes a coenzyme B12- dependent deamination of vicinal amino alcohols. The mode of binding of coenzyme B12 to EAL has been investigated by electron paramagnetic resonance spectroscopy (EPR) using [15N]-dimethylbenzimidazole-coenzyme B12. EAL was incubated with either unlabeled or 15N-enriched coenzyme B12 and then either exposed to light or treated with ethanol to generate the cleaved form of the cofactor, cob(II)alamin (B12(r)) bound in the active site. The reaction mixtures were examined by EPR spectroscopy at 77 K. 15N superhyperfine splitting in the EPR signals of the low-spin Co2+ of B12(r), bound in the active site of EAL, indicates that the dimethylbenzimidazole moiety of the cofactor contributes the lower axial ligand consistent with 'base-on' binding of coenzyme B12 to EAL.

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DO - 10.1006/abbi.1999.1382

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JO - Archives of Biochemistry and Biophysics

JF - Archives of Biochemistry and Biophysics

IS - 1

ER -

Ethanolamine ammonia-lyase has a 'base-on' binding mode for coenzyme B₁₂

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