1992 Fiscal Year Final Research Report Summary
Development of Crystal Structure Analysis Methods for Small Protein Crystals Using Anomalous Scattering.
| Project/Area Number |
02558011
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| Research Category |
Grant-in-Aid for Developmental Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
結晶学
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| Research Institution | UNIVERSITY OF TOKYO |
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Principal Investigator |
SATOW Yoshinori University of Tokyo, Faculty of Pharmaceutical Sciences. Professor, 薬学部, 教授 (30150014)
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| Co-Investigator(Kenkyū-buntansha) |
NOGUCHI Shuji University of Tokyo, Faculty of Pharmaceutical Sciences. Assistant, 薬学部, 助手 (60237823)
NAKASAKO Masayoshi University of Tokyo, Faculty of Pharmaceutical Sciences. Assistant, 薬学部, 助手 (30227764)
AMEMIYA Yoshiyuki National Laboratory for High Energy Physics, Photon Factory. Associate Professor, 放射光実験施設, 助教授 (70151131)
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| Project Period (FY) |
1990 – 1992
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| Keywords | Small Crystal / Protein / X-Ray Crystallography / Imaging Plate / Synchrotron Radiation / Anomalous Scattering / Immunoglobulin / Antibody |
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| Research Abstract |
The research objective is to develop an X-ray area detector system, utilizing Imaging Plate storage phosphor, for structural studies of small crystals of protein. This study aimed at development of structure determination theory and data processing methods by use of anomalous scattering. Computer programs developed here, as well as other existing structure refinements and graphics programs, were built into a complete system for anomalous diffraction studies.
A drum-scanner type X-ray detector was designed and built first. The detector implemented with highly efficient optical system of new design was combined with an Enraf-Nonius oscillation camera for a rotating anode X-ray generator. The detector can be used in oscillation data collection when it is mounted on the horizontal-type four-circle diffractometer system installed at beamline 14A of the Photo Factory synchrotron radiation source at National Laboratory for High Energy Physics. A preliminary application of the detector and program system to a Laue diffraction study was perfomed on the hen egg white lysozyme in order to evaluate feasibility of white radiation anomalous diffraction.
Three protein structures of very small crystals were determined based on the synchrotron data measured at several wavelengths. The structure of an Fv fragment from mouse anti-dansyllysine immunoglobulin IgG2a has been determined, finally at 1.6 A^^. resolution. The structure clearly elucidates its complementarity determining regions. Refinement of its crystalline complex with dasyllysine are also in progress. Furthermore, crystal structures of purine-specific ribonuclease U_2 at 1.8 A^^. resolution and of an Fab fragment of 3B44 anti-nitrophenol antibody of IgG12 at 3 A^^. have been determined. These have proved that the developed detector and the program system are widely applicable to structure determination of protein crystals as well as 50 mum.
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