Photolysis of the Carbon Monoxide Complex of Myoglobin: Nanosecond Time-Resolved Crystallography

The biological activity of macromolecules is accompanied by rapid structural changes. The photosensitivity of the carbon monoxide complex of myoglobin was used at the European Synchrotron Radiation Facility to obtain pulsed, Laue x-ray diffraction data with nanosecond time resolution during the process of heme and protein relaxation after carbon monoxide photodissociation and during rebinding. These time-resolved experiments reveal the structures of myoglobin photoproducts, provide a structural foundation to spectroscopic results and molecular dynamics calculations, and demonstrate that time-resolved macromolecular crystallography can elucidate the structural bases of biochemical mechanisms on the nanosecond time scale.

[1]  R. Miller,et al.  Direct observation of global protein motion in hemoglobin and myoglobin on picosecond time scales. , 1991, Science.

[2]  M. Lim,et al.  Binding of CO to myoglobin from a heme pocket docking site to form nearly linear Fe-C-O , 1995, Science.

[3]  K. Moffat,et al.  Feasibility and Realization of Single-Pulse Laue Diffraction on Macromolecular Crystals at ESRF. , 1996, Journal of synchrotron radiation.

[4]  M. Karplus,et al.  Nonexponential relaxation after ligand dissociation from myoglobin: a molecular dynamics simulation. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[5]  Q. Gibson,et al.  A kinetic description of ligand binding to sperm whale myoglobin. , 1986, The Journal of biological chemistry.

[6]  P. Champion,et al.  Probing picosecond processes with nanosecond lasers: Electronic and vibrational relaxation dynamics of heme proteins , 1992 .

[7]  R. Hochstrasser,et al.  Picosecond transient absorption study of photodissociated carboxy hemoglobin and myoglobin. , 1988, Biophysical journal.

[8]  K. D. Straub,et al.  Direct observation of hot vibrations in photoexcited deoxyhemoglobin using picosecond Raman spectroscopy , 1991 .

[9]  Schmitt,et al.  Empirical determination of universal multifractal exponents in turbulent velocity fields. , 1992, Physical review letters.

[10]  D E Koshland,et al.  Mutagenesis and Laue structures of enzyme intermediates: isocitrate dehydrogenase. , 1995, Science.

[11]  S. Franzen,et al.  Evidence for sub-picosecond heme doming in hemoglobin and myoglobin: a time-resolved resonance Raman comparison of carbonmonoxy and deoxy species. , 1995, Biochemistry.

[12]  K. Moffat,et al.  Optical studies of a bacterial photoreceptor protein, photoactive yellow protein, in single crystals. , 1995, Biochemistry.

[13]  M. Lim,et al.  Mid-infrared vibrational spectrum of CO after photodissociation from heme: Evidence for a ligand docking site in the heme pocket of hemoglobin and myoglobin , 1995 .

[14]  Z. Ren,et al.  Quantitative Analysis of Synchrotron Laue Diffraction Patterns in Macromolecular Crystallography , 1995 .

[15]  K. Moffat,et al.  Optical monitoring of protein crystals in time‐resolved x‐ray experiments: Microspectrophotometer design and performance , 1994 .

[16]  J. Simon,et al.  Protein conformational relaxation following photodissociation of CO from carbonmonoxymyoglobin: picosecond circular dichroism and absorption studies. , 1991, Biochemistry.

[17]  R. Henderson,et al.  Freeze trapping of reaction intermediates. , 1995, Current opinion in structural biology.

[18]  D. Morikis,et al.  Conformational interconversion in protein crystals. , 1992, Journal of molecular biology.

[19]  C. M. Jones,et al.  Conformational relaxation and ligand binding in myoglobin. , 1994, Biochemistry.

[20]  M. Lim,et al.  Nonexponential protein relaxation: dynamics of conformational change in myoglobin. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[21]  K. Moffat Time-resolved macromolecular crystallography , 1996 .

[22]  J. Straub,et al.  Molecular dynamics simulation of NO recombination to myoglobin mutants. , 1993, The Journal of biological chemistry.

[23]  D. Ringe,et al.  Charge motion in MbCO crystals after flash photolysis. , 1989, Biophysical journal.

[24]  Champion,et al.  Relaxation dynamics of myoglobin in solution. , 1992, Physical review letters.

[25]  R. Hochstrasser,et al.  Molecular dynamics simulations of cooling in laser-excited heme proteins. , 1986, Proceedings of the National Academy of Sciences of the United States of America.

[26]  K. Moffat,et al.  Photolysis-induced structural changes in single crystals of carbonmonoxy myoglobin at 40 K , 1994, Nature Structural Biology.

[27]  G. Nienhaus,et al.  X-ray structure determination of a metastable state of carbonmonoxy myoglobin after photodissociation. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[28]  K. Moffat,et al.  X-ray Laue Diffraction from Protein Crystals , 1984, Science.

[29]  R. Riopelle,et al.  Survival of Cholinergic Forebrain Neurons in Developing p75NGFR-Deficient Mice , 1996, Science.

[30]  M Karplus,et al.  X-ray structure and refinement of carbon-monoxy (Fe II)-myoglobin at 1.5 A resolution. , 1986, Journal of molecular biology.

[31]  E. Henry,et al.  Simulation of the kinetics of ligand binding to a protein by molecular dynamics: geminate rebinding of nitric oxide to myoglobin. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[32]  M. Karplus,et al.  Enhanced sampling in molecular dynamics: use of the time-dependent Hartree approximation for a simulation of carbon monoxide diffusion through myoglobin , 1990 .

[33]  H Frauenfelder,et al.  Dynamics of ligand binding to myoglobin. , 1975, Biochemistry.