Refined crystal structure of Streptomyces griseus trypsin at 1.7 A resolution.

Abstract

Streptomyces griseus trypsin (SGT) is a bacterial serine proteinase that is more homologous to mammalian than to other bacterial enzymes. The structure of SGT has been solved primarily by molecular replacement, though some low-resolution phase information was supplied by heavy-atom derivatives. The mammalian pancreatic serine proteinases bovine trypsin (BT) and alpha-chymotrypsin (CHT) were used as molecular replacement models. Because these proteins have low homology with SGT compared to the majority of other successful replacement models, new strategies were required for molecular replacement to succeed. The model of SGT has been refined at 1.7 A resolution to a final R-factor of 0.161 (1 A = 0.1 nm); the correlation coefficient between all observed and calculated structure factor amplitudes is 0.908. Solvent molecules located in the crystal structure play an important role in stabilizing buried charged and polar groups. An additional contribution to stability can be seen in the fact that the majority of the charged side-chains are involved in ionic interactions, sometimes linking the two domains of SGT. A comparison of SGT with BT shows that the greatest similarities are in the active-site and substrate-binding regions, consistent with their similar substrate specificities. The modeling of complexes of SGT with two inhibitors of BT, pancreatic trypsin inhibitor (PTI) and the third domain of Japanese quail ovomucoid (OMJPQ3), helps to explain why PTI inhibits SGT but OMJPQ3 does not. Like BT, but unlike other bacterial serine proteinases of known structure, SGT has a buried N terminus. SGT has also a well-defined Ca2+-binding site, but this site differs in location from that of BT.