Crystal Structure of Streptomyces erythraeus Trypsin at 2.7 Å Resolution

Abstract

The crystal structure of Streptomyces erythraeus trypsin (abbreviated as SET) has been determined in order to clarify the precise structure of the vicinity of the active site of serine protease and to understand its structure-function relationship. Crystals of SET were prepared at its active pH range (pH 5-10) without any inhibitors which might have affected the circumstances around the active sites. The structure model of SET was made based on the electron density map obtained by the multiple isomorphous replacement method at 3.5 Å resolution, and refined by the restrained least-squares method. The current model yields a crystallographic R-factor of 0.272 for 4, 968 reflections between 8 and 2.7 Å resolution. Though the sequence homology among SET, Streptomyces griseus trypsin and bovine trypsin, 32-37%, is not so high, their overall structures are similar to each other. Compari-son of the three molecular structures shows that: 1) the folding of the main chains of the three proteins is essentially the same though there are significant differences on the molecular surface; 2) the spatial arrangements of the catalytic triads in the three proteins are similar to each other; 3) in SET and S. griseus trypsin a short stretch of 3₁₀-helix is found through A1a56 to Thr59; His57 in this segment is one important amino acid residue involved in the active sites.