Single-crystal EPR and ENDOR study of nitrogenase from clostridium pasteurianum

Ryszard J. Gurbiel*, Jeffrey T. Bolin, Alicia E. Ronco, Leonard Mortenson, Brian M. Hoffman

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

14 Scopus citations

Abstract

The molybdenum-iron (MoFe) protein of nitrogenase from Clostridium pasteurianum forms monoclinic crystals in space group P21. The unit cell comprises two molecules, each of which possesses a molecular twofold symmetry axis and binds two chemically identical molybdenum-iron cofactors (M centers). Thus, there are four magnetically distinct M centers within the unit cell related by two different symmetry operations: a molecular twofold axis of symmetry in the bc* plane relates the two EPR centers within a molecule; a crystallographic twofold screw axis of symmetry along the b axis of the unit cell relates the two different molecules in a unit cell. Single-crystal EPR studies at X-band (9.5 GHz) and Q-band (35 GHz) microwave frequencies have been employed to determine g tensors in the crystallographic frame for the four magnetically distinct M centers. Determination of these tensors has been achieved by a novel procedure that relies heavily on the symmetry relations between sites; it rests primarily on measurements that only involve rotation of the magnetic field parallel to a single plane of the crystal and uses g values from frozen-solution samples instead of additional rotations. Single-crystal 1H ENDOR spectra of the M centers are presented.

Original languageEnglish (US)
Pages (from-to)227-240
Number of pages14
JournalJournal of Magnetic Resonance (1969)
Volume91
Issue number2
DOIs
StatePublished - Feb 1 1991

Funding

This work has been supported by the USDA (87~CRCR-1-2430 BMH; 87-CRCR-I-2429 JTB), the NSF (DMB 8907559 BMH), and the NIH (GM40067 LM, and also HL13531 BMH). Its inception owes a great debt to the insights and encouragement of Professor W. H. Orme-Johnson. We acknowledge the invaluable technical efforts of Clark Davoust.

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