Contaminant inclusion into protein crystals analyzed by electrospray mass spectrometry and X-ray crystallography.
The inclusion of protein contaminants into crystals of turkey egg white lysozyme (TEWL) was investigated by electrospray mass spectrometry of the dissolved crystals. The results show that significant amounts of the structurally related contaminant hen egg white lysozyme (HEWL) are included in the crystals of TEWL. The structurally unrelated contaminant RNAse A, on the other hand, is not included. The X-ray diffraction data statistics of a hybrid TEWL/HEWL crystal and an uncontaminated TEWL crystal were of similar quality. This indicates that, even though the crystals contain much higher levels of the contaminant than one would have expected after a recrystallization experiment, they are still suitable for X-ray diffraction experiments. However, attempts to detect the presence of the contaminant in the crystal by crystallographic structure refinement did not yield conclusive results.
Hirschler
J
J
Laboratoire de Cristallographie et de Cristallogenèse des Protéines, Institut de Biologie Structurale J.-P. Ebel (CEA-CNRS) 41, Grenoble, France.
Halgand
F
F
Forest
E
E
Fontecilla-Camps
J C
JC
D000818:Animals; D002645:Chickens; D003460:Crystallization; D018360:Crystallography, X-Ray; D004529:Egg White; D013058:Mass Spectrometry; D008958:Models, Molecular; D009113:Muramidase / Q000032:analysis / Q000737:chemistry*; D012259:Ribonuclease, Pancreatic / Q000737:chemistry; D014422:Turkeys
D012259:Ribonuclease, Pancreatic; D009113:Muramidase