Analyzing protein functions in four dimensions.
Time-resolved structural studies on biomolecular function are coming of age. Focus has shifted from studies on 'systems of opportunities' to a more problem-oriented approach, addressing significant questions in biology and chemistry. An important step in this direction has been the use of physical and chemical trapping methods to capture and then freeze reaction intermediates in crystals. Subsequent monochromatic data collection at cryogenic temperatures can produce high resolution structures of otherwise elusive intermediates. The combination of diffraction methods with spectroscopic techniques provides a means to directly correlate electronic transitions with structural transitions in the sample, eliminating much of the guesswork from experiments. Studies on cytochrome P450, isopenicillin N synthase, cytochrome cd1 nitrite reductase, copper amine oxidase and bacteriorhodopsin were selected as examples, and the results are discussed.
Hajdu
J
J
Department of Biochemistry, Uppsala University, Biomedical Center, Box 576, S-751 23 Uppsala, Sweden. janos@xray.bmc.uu.se
Neutze
R
R
Sjögren
T
T
Edman
K
K
Szöke
A
A
Wilmouth
R C
RC
Wilmot
C M
CM
D006631:Amine Oxidase (Copper-Containing) / Q000737:chemistry / Q000378:metabolism; D001436:Bacteriorhodopsins / Q000737:chemistry / Q000378:metabolism; D003460:Crystallization; D018360:Crystallography, X-Ray / Q000379:methods*; D003574:Cytochrome c Group; D003580:Cytochromes / Q000737:chemistry / Q000378:metabolism; D004058:Diffusion; D004789:Enzyme Activation; D008958:Models, Molecular; D009572:Nitrite Reductases / Q000737:chemistry / Q000378:metabolism; D010088:Oxidoreductases / Q000737:chemistry / Q000378:metabolism; D010100:Oxygen / Q000378:metabolism; D011487:Protein Conformation; D011506:Proteins / Q000737:chemistry* / Q000378:metabolism*; D013997:Time Factors
D003574:Cytochrome c Group; D003580:Cytochromes; D011506:Proteins; D001436:Bacteriorhodopsins; C019315:cytochrome cd1; D010088:Oxidoreductases; C038370:isopenicillin N synthetase; D006631:Amine Oxidase (Copper-Containing); D009572:Nitrite Reductases; D010100:Oxygen