Characterization of crystal content by ESI-MS and MALDI-MS.
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).
Potier
N
N
Laboratoire de Spectrométrie de Masse Bio-Organique, 1 Rue Blaise Pascal, 67008 Strasbourg, France.
Lamour
V
V
Poterszman
A
A
Thierry
J C
JC
Moras
D
D
Van Dorsselaer
A
A
D000449:Aldehyde Reductase / Q000737:chemistry; D003460:Crystallization; D004268:DNA-Binding Proteins / Q000737:chemistry; D008024:Ligands; D009113:Muramidase / Q000737:chemistry*; D009249:NADP / Q000737:chemistry; D009841:Oligonucleotides / Q000737:chemistry; D018160:Receptors, Cytoplasmic and Nuclear / Q000737:chemistry; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D035181:TATA-Box Binding Protein; D014157:Transcription Factors / Q000737:chemistry
D004268:DNA-Binding Proteins; D008024:Ligands; D009841:Oligonucleotides; D018160:Receptors, Cytoplasmic and Nuclear; D035181:TATA-Box Binding Protein; D014157:Transcription Factors; D009249:NADP; D000449:Aldehyde Reductase; D009113:Muramidase