期刊
JOURNAL OF APPLIED CRYSTALLOGRAPHY
卷 49, 期 -, 页码 149-157出版社
INT UNION CRYSTALLOGRAPHY
DOI: 10.1107/S1600576715023195
关键词
high-pressure cryocooling; protein crystal disorder; mosaicity; nonmerohedral twinning
资金
- National Science Foundation
- National Institutes of Health/National Institute of General Medical Sciences under NSF [DMR-1332208]
- National Institute of General Medical Sciences, National Institutes of Health [GM-103485]
High-pressure cryocooling (HPC) has been developed as a technique for reducing the damage that frequently occurs when macromolecular crystals are cryocooled at ambient pressure. Crystals are typically pressurized at around 200 MPa and then cooled to liquid nitrogen temperature under pressure; this process reduces the need for penetrating cryoprotectants, as well as the damage due to cryocooling, but does not improve the diffraction quality of the as-grown crystals. Here it is reported that HPC using a pressure above 300 MPa can reduce lattice disorder, in the form of high mosaicity and/or nonmerohedral twinning, in crystals of three different proteins, namely human glutaminase C, the GTP pyrophosphokinase YjbM and the uncharacterized protein lpg1496. Pressure lower than 250 MPa does not induce this transformation, even with a prolonged pressurization time. These results indicate that HPC at elevated pressures can be a useful tool for improving crystal packing and hence the quality of the diffraction data collected from pressurized crystals.
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