Specific chemical and structural damage to proteins produced by synchrotron radiation.
Radiation damage is an inherent problem in x-ray crystallography. It usually is presumed to be nonspecific and manifested as a gradual decay in the overall quality of data obtained for a given crystal as data collection proceeds. Based on third-generation synchrotron x-ray data, collected at cryogenic temperatures, we show for the enzymes Torpedo californica acetylcholinesterase and hen egg white lysozyme that synchrotron radiation also can cause highly specific damage. Disulfide bridges break, and carboxyl groups of acidic residues lose their definition. Highly exposed carboxyls, and those in the active site of both enzymes, appear particularly susceptible. The catalytic triad residue, His-440, in acetylcholinesterase, also appears to be much more sensitive to radiation damage than other histidine residues. Our findings have direct practical implications for routine x-ray data collection at high-energy synchrotron sources. Furthermore, they provide a direct approach for studying the radiation chemistry of proteins and nucleic acids at a detailed, structural level and also may yield information concerning putative "weak links" in a given biological macromolecule, which may be of structural and functional significance.
Weik
M
M
Department of Crystal and Structural Chemistry, Bijvoet Center for Biomolecular Research, Utrecht University, Utrecht, NL-3584 CH, The Netherlands.
Ravelli
R B
RB
Kryger
G
G
McSweeney
S
S
Raves
M L
ML
Harel
M
M
Gros
P
P
Silman
I
I
Kroon
J
J
Sussman
J L
JL
D000110:Acetylcholinesterase / Q000737:chemistry / Q000528:radiation effects; D000818:Animals; D002645:Chickens; D003460:Crystallization; D018360:Crystallography, X-Ray; D004220:Disulfides / Q000737:chemistry / Q000528:radiation effects; D004529:Egg White; D009113:Muramidase / Q000737:chemistry / Q000528:radiation effects; D011487:Protein Conformation / Q000528:radiation effects; D011506:Proteins / Q000737:chemistry* / Q000528:radiation effects*; D011829:Radiation Dosage; D017356:Synchrotrons; D014101:Torpedo
D004220:Disulfides; D011506:Proteins; D000110:Acetylcholinesterase; D009113:Muramidase