Structures from anomalous diffraction of native biological macromolecules.
Crystal structure analyses for biological macromolecules without known structural relatives entail solving the crystallographic phase problem. Typical de novo phase evaluations depend on incorporating heavier atoms than those found natively; most commonly, multi- or single-wavelength anomalous diffraction (MAD or SAD) experiments exploit selenomethionyl proteins. Here, we realize routine structure determination using intrinsic anomalous scattering from native macromolecules. We devised robust procedures for enhancing the signal-to-noise ratio in the slight anomalous scattering from generic native structures by combining data measured from multiple crystals at lower-than-usual x-ray energy. Using this multicrystal SAD method (5 to 13 equivalent crystals), we determined structures at modest resolution (2.8 to 2.3 angstroms) for native proteins varying in size (127 to 1148 unique residues) and number of sulfur sites (3 to 28). With no requirement for heavy-atom incorporation, such experiments provide an attractive alternative to selenomethionyl SAD experiments.
New York Structural Biology Center, National Synchrotron Light Source (NSLS) X4, Brookhaven National Laboratory, Upton, NY 11973, USA.
Qun
Q
Liu
Tassadite
T
Dahmane
Zhen
Z
Zhang
Zahra
Z
Assur
Julia
J
Brasch
Lawrence
L
Shapiro
Filippo
F
Mancia
Wayne A
WA
Hendrickson
D001426:Bacterial Proteins / Q000737:chemistry; D018360:Crystallography, X-Ray / Q000379:methods*; D003627:Data Interpretation, Statistical; D058851:GPI-Linked Proteins / Q000737:chemistry; D000071677:Histidine Kinase; D008958:Models, Molecular; D009419:Nerve Tissue Proteins / Q000737:chemistry; D000075385:Netrins; D011487:Protein Conformation; D011494:Protein Kinases / Q000737:chemistry; D017434:Protein Structure, Tertiary; D011506:Proteins / Q000737:chemistry*; D012645:Selenomethionine / Q000737:chemistry; D059629:Signal-To-Noise Ratio; D013455:Sulfur / Q000737:chemistry; D014961:X-Ray Diffraction
D001426:Bacterial Proteins; D058851:GPI-Linked Proteins; C463083:NTNG2 protein, human; D009419:Nerve Tissue Proteins; D000075385:Netrins; D011506:Proteins; D013455:Sulfur; D012645:Selenomethionine; D011494:Protein Kinases; D000071677:Histidine Kinase