Crystallization and preliminary X-ray crystallographic analysis of the complex between the N-D1 domain of VCP from Homo sapiens and the N domain of OTU1 from Saccharomyces cerevisiae

Abstract

VCP (valosin-containing protein; also known as p97) plays important roles in many biological processes including the ERAD (endoplasmic reticulum-associated degradation) pathway and its function is governed by binding partners. OTU1 (ovarian tumour domain-containing protein 1) is a recently discovered deubiquitinating enzyme that interacts directly with VCP in the ERAD pathway. In order to understand the interactions between the two proteins, the N-D1 domain of VCP and the UBXL domain of OTU1 were cloned, overexpressed, purified and crystallized. The crystals of the complex diffracted to 3.25 angstrom resolution and belonged to space group P2(1), with unit-cell parameters a = 165.45, b = 176.73, c = 165.59 angstrom, beta = 120.095 degrees. There are two molecules of the complex in the asymmetric unit with a Matthews coefficient of 2.62 angstrom(3) Da(-1) and a solvent content of 53%.