Open AccessA new crystal form of human vascular adhesion protein 1
Author(s) -
Ernberg Karin,
McGrath Aaron P.,
Peat Thomas S.,
Adams Timothy E.,
Xiao Xiaowen,
Pham Tam,
Newman Janet,
McDonald Ian A.,
Collyer Charles A.,
Guss J. Mitchell
Publication year - 2010
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309110041515
Subject(s) - chinese hamster ovary cell , adhesion , glycosylation , chemistry , crystallography , quinone , molecule , cell adhesion , tetramer , stereochemistry , biochemistry , cell , enzyme , receptor , organic chemistry
Human vascular adhesion protein 1 (VAP‐1) is involved in lymphocyte–endothelial cell adhesion and has been implicated in many human inflammatory diseases. VAP‐1 is a member of the copper amine oxidase family of enzymes with a trihydroxyphenylalanine quinone (TPQ) cofactor. Previously characterized crystals of VAP‐1 suffered from anisotropy and contained disordered regions; in addition, one form was consistently twinned. In an effort to grow crystals that diffracted to higher resolution for inhibitor‐binding studies, a construct with an N‐terminal deletion was made and expressed in the Chinese hamster ovary (CHO) glycosylation mutant cell line Lec8. Screening produced crystals that displayed some anisotropy and contained seven molecules per asymmetric unit. These crystals belonged to space group C 2, with unit‐cell parameters a = 394.5, b = 115.8, c = 179.3 Å, β = 112.3°. The structure was refined to a resolution of 2.9 Å, with R cryst and R free values of 0.250 and 0.286, respectively.