Secondary structures and structural fluctuation in a dimeric protein, Streptomyces subtilisin inhibitor.
Based on the nuclear magnetic resonance assignments of a dimeric protein, Streptomyces subtilisin inhibitor (SSI), microscopic details of secondary structures in solution have been elucidated. The chemical shift index of C(alpha) signals, together with information on the hydrogen exchange rates of the backbone amide protons, were used to identify secondary structures. The locations of these secondary structures were found to be different in some critical points from those determined earlier by X-ray crystallography of the crystal. Notably, the beta3 strand is completely missing and the alpha2 helix is extended toward the C-terminus. Furthermore, hydrogen exchange experiments of individual peptide NH protons under strongly folding conditions revealed mechanisms of global and local structural fluctuation within the dimeric structure. It has been suggested that the global fluctuation of the monomeric unit occurs without affecting the accompanying monomer, in contrast to the equilibrium thermal unfolding, which is cooperative. Higher protection against hydrogen exchange for residues in part of the beta4 strand implies that this region might serve as a folding core.
Sasakawa
H
H
Graduate School of Science and Technology, Kobe University, Nada-ku, Kobe, 657-8501, Japan.
Tamura
A
A
Fujimaki
S
S
Taguchi
S
S
Akasaka
K
K
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry*; D019281:Dimerization; D009682:Magnetic Resonance Spectroscopy; D008958:Models, Molecular; D020836:Protein Structure, Quaternary; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D015842:Serine Proteinase Inhibitors / Q000737:chemistry*; D013302:Streptomyces / Q000737:chemistry*; D013381:Subtilisins / Q000037:antagonists & inhibitors*; D013816:Thermodynamics
D001426:Bacterial Proteins; D015842:Serine Proteinase Inhibitors; C018150:subtilisin inhibitor protein, Streptomyces; D013381:Subtilisins