Distinct Structural Features of The A and β Subunits of Nitrogenase Molybdenum-iron Protein of Clostridium Pasteurianum: An Analysis of Amino Acid Sequences

Academic Article

Abstract

  • Nitrogenase is composed of two separately purified proteins, a molybdenum-iron (MoFe) protein and an iron (Fe) protein. Structural genes (nifD and nifK) encoding a and β subunits of the MoFe protein of Clostridium pasteurianum (Cp) have been cloned and sequenced. The deduced amino acid sequences were analyzed for structures that could be related to the unique properties of the Cp protein, particularly its low capacity to form an active enzyme with a heterologous Fe protein. Cp nifK is located immediately downstream from Cp nifD, with the start codon of nifK overlapping by one base with the stop codon of nifD. An open reading frame following nifK was identified as nifE. The amino acid sequence deduced from nifK encompasses the partial amino acid sequences previously reported from the isolated β subunit. Cp nifK encodes a polypeptide of 458 amino acid residues (Mr50115) whose amino-terminal region is about 50 residues shorter than the otherwise conserved corresponding polypeptides from four other organisms. In contrast, Cp a subunit (nifD product) contains an additional stretch of 50 amino acid residues in the 380–430 region, which is unique to the Cp protein. It therefore appears that the combined size of the a and β subunits could be important to nitrogenase function. An analysis of the predicted secondary structure from the amino acid sequence of each subunit from three species (C. pasteurianum, Azotobacter vinelandii, and Rhizobium japonicum) further revealed structural features, including regions adjacent to some of the conserved cysteine residues, differentiating the Cp MoFe protein from others. These different regions may be further tested for correlation with distinct properties of Cp nitrogenase. © 1988, American Chemical Society. All rights reserved.
  • Authors

    Digital Object Identifier (doi)

    Author List

  • Wang SZ; Chen JS; Johnson L
  • Start Page

  • 2800
  • End Page

  • 2810
  • Volume

  • 27
  • Issue

  • 8