The primary structure of extracellular Bacillus polymyxa ribonuclease (RNase Bpo) was established by mass spectroscopy analysis and automatic Edman degradation of the individual peptides obtained from protein digestion with Glu-specific protease V8. RNase Bpo consists of 111 amino acid residues, with a relative molecular weight of 12,607. RNase Bpo is a close structural homolog of RNases of B. amyloliquefaciens (RNase Ba) and B. intermedius (RNase Bi), the similarity of their primary structures being 68%. Molecular modelling of the structure of the complex of RNase Bpo with substrate analog d(CGAC) was performed and a spatial model based on the known crystal structure of RNase Ba complex with the corresponding nucleotide was constructed using the methods of interactive computer graphics and energy minimization. The differences in the primary and tertiary structures of the enzymes were analyzed in order to understand the substrate specificity of Bacillus RNases.
Lebedev, A. A., Shlyapnikov, S. V., Pustobaev, V. N., Kirpichnikov, M. P., & Dementiev, A. A. (1996). Structural characterization of extracellular ribonuclease of Bacillus polymyxa: Amino acid sequence determination and spatial structure prediction. FEBS Letters, 392(2), 105–109. https://doi.org/10.1016/0014-5793(96)00793-4