Characterization of crystal content by ESI-MS and MALDI-MS.
Fiche publication
Date publication
décembre 2000
Auteurs
Membres identifiés du Cancéropôle Est :
Dr MORAS Dino, Dr POTERSZMAN Arnaud, Dr VAN DORSSELAER Alain
Tous les auteurs :
Potier N, Lamour V, Poterszman A, Thierry JC, Moras D, Van Dorsselaer A
Lien Pubmed
Résumé
A general approach based on mass spectrometry is described for the rapid identification of the content of macromolecular crystals. The experimental procedure was established using lysozyme crystals and then successfully applied to various systems containing specifically bound molecules not easily detectable by other classical techniques. This procedure can be carried out on crystals containing macromolecules of a different nature, such as proteins, nucleic acids and small organic molecules and their non-covalent complexes, grown under various crystallization conditions including PEGs and salts. It can be applied very early on in the crystallization process - as soon as the crystals can be handled. It allows the biologist to control precisely the sequence integrity and homogeneity of the crystallized proteins (in particular at the C-terminus) as well as to verify whether the protein has crystallized with all its expected partners or ligands (nucleic acid molecules, cofactor or small organic molecules).
Référence
Acta Crystallogr D Biol Crystallogr. 2000 Dec;56(Pt 12):1583-90.