H n.m.r. spectrum from the flexible N-terminal segment of Streptomyces subtilisin inhibitor.
The 1H n.m.r. spectrum of Streptomyces subtilisin inhibitor shows a limited number of unusually sharp signals at room temperature. Some of these signals are assigned uniquely to protons of the side chains of the N-terminal segment, Aspl-Ala2-Pro3-Ser4-Ala5-Leu6-Tyr7-based on experiments of spin decoupling, pH titration, and enzymatic cleavage of the protein. Quantitative examination of these signals indicates that the N-terminal end of this protein is heterogeneous in that the protein contains a considerable fraction whose sequence starts with Ala2 rather than with Asp1. The pKa values for the amino groups of Asp1 and Ala2 exposed at the N-terminus are determined to be 8.9 +/- 0.4 and 9.0 +/- 0.1, respectively. Furthermore, examination of the line-widths of the methyl proton resonances of Ala2 and Ala5 residues indicates that the N-terminal peptide segment is free and undergoes rapid segmental motions in the order of 10(-9) s.
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000302:isolation & purification; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D011487:Protein Conformation; D013302:Streptomyces; D013381:Subtilisins / Q000037:antagonists & inhibitors*
D001426:Bacterial Proteins; C018150:subtilisin inhibitor protein, Streptomyces; D013381:Subtilisins