X-ray scattering using synchrotron radiation shows nitrite reductase from Achromobacter xylosoxidans to be a trimer in solution.
We demonstrate here the applicability of X-ray scattering for studying molecular conformation of multimeric proteins in solution by using synchrotron radiation to extend the range of data collection to include medium angles (ca. 3-4 degrees). We have been able to define the solution structure of the dissimilatory nitrite reductase of Achromobacter xylosoxidans (AxNiR), an enzyme for which there are conflicting reports as to the nature of its multimeric structure. Quantitative interpretation of the X-ray scattering profile, based on a modeling study using the high-resolution crystal structure data for the nitrite reductase from the related organism Achromobacter cycloclastes (AcNiR), provides a detailed model for the trimeric structure of AxNiR in solution. Sedimentation equilibrium centrifugation gave an M(r) of 103,000, consistent with such a trimeric structure.
Grossmann
J G
JG
Molecular Biophysics Group, SERC Daresbury Laboratory, Warrington, Cheshire, U.K.
Abraham
Z H
ZH
Adman
E T
ET
Neu
M
M
Eady
R R
RR
Smith
B E
BE
Hasnain
S S
SS
D000421:Alcaligenes / Q000201:enzymology*; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D003460:Crystallization; D046911:Macromolecular Substances; D008433:Mathematics; D008958:Models, Molecular; D008970:Molecular Weight; D009572:Nitrite Reductases / Q000737:chemistry*; D011487:Protein Conformation; D012542:Scattering, Radiation; D012996:Solutions; D017356:Synchrotrons; D014461:Ultracentrifugation; D014965:X-Rays
D046911:Macromolecular Substances; D012996:Solutions; D009572:Nitrite Reductases