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Characterization of a FeMo cofactor-deficient MoFe protein from anifE-deleted strain (DJ35) ofAzotobacter vinelandii

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Chinese Science Bulletin

Abstract

A MoFe protein (ΔnifE Avl) with a purity of ∼80% was purified from a nifE-deleted mutant ofAzotobacter vinelandii DJ35. Compared with MoFe protein purified from wild-type strain OP (OP Av1), ΔnifE Av1 had the same subunits composition, and had immune reaction with antibody to OP Av1, but its relative mobility in anaerobic native polyacrylamide gel electrophoresis (PAGE) was a little larger than that of OP Av1. Metal analysis showed that Mo and Fe contents of ΔnifE Av1 both apparently decreased. When complemented with OP Fe protein, ΔnifE Av1 had no C2H2-reduction activity, but it could bein vitro activated by FeMoco extracted from OP Av1. The circular dichroism (CD) spectrum of ΔnifE Av1 at ∼450 nm was similar to that of OP Av1, while the EPR signal at g∼3.7 was absolutely silent, and the signal intensities at g∼=4.3 and 2.0 decreased by 75% and 50%, respectively. The results indicated that ΔnifE Av1 purified from DJ35 was a FeMoco-deficient but P-cluster-containing MoFe protein.

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Authors and Affiliations

  1. Key Laboratory of Photosynthesis and Environmental Molecular Physiology, Institute of Botany, Chinese Academy of Sciences, 100093, Beijing, China

    Ying Zhao, Shaomin Bian, Huina Zhou, Huangping Wang, Jianfeng Zhao & Jufu Huang

  2. Graduate School, Chinese Academy of Sciences, 100039, Beijing, China

    Ying Zhao, Shaomin Bian & Huina Zhou

  3. Key Laboratory of Photochemistry, Institute of Chemistry, Chinese Academy of Sciences, 100080, Beijing, China

    Chunxi Zhang

  4. Bioengineering College, Fujian Normal University, 360007, Fuzhou, China

    Huangping Wang

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  1. Ying Zhao
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  2. Shaomin Bian
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  3. Chunxi Zhang
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  4. Huina Zhou
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  6. Jianfeng Zhao
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  7. Jufu Huang
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Correspondence to Jufu Huang.

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Zhao, Y., Bian, S., Zhang, C. et al. Characterization of a FeMo cofactor-deficient MoFe protein from anifE-deleted strain (DJ35) ofAzotobacter vinelandii . Chin.Sci.Bull. 50, 2305–2310 (2005). https://doi.org/10.1007/BF03183740

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  • DOI: https://doi.org/10.1007/BF03183740

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