Open AccessEukaryotic expression, purification, crystallization and preliminary X‐ray analysis of murine Manic Fringe
Author(s) -
Jinek Martin,
Conti Elena
Publication year - 2006
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s174430910602611x
Subject(s) - crystallization , materials science , crystallography , microbiology and biotechnology , chemistry , biology , organic chemistry
Fringe proteins are Golgi‐resident β1,3‐ N ‐acetylglucosaminyltransferases that regulate development in metazoa through glycosylation of the Notch receptor and its ligands. The catalytic domain of murine Manic Fringe was expressed in the baculovirus/insect‐cell system as a secreted protein. Mass‐spectrometric analysis of the purified protein indicated the presence of two N‐linked glycans. Abolishing the glycosylation sites by site‐directed mutagenesis was necessary in order to obtain orthorhombic crystals that diffracted to 1.8 Å resolution. For phasing, a highly redundant data set was collected using a crystal soaked with halide salts.