An Unexpected Outcome of Surface Engineering An Integral Membrane Protein: Improved Crystallization of Cytochrome Ba(3) From Thermus Thermophilus
Journal Article
·
· Acta Crystallogr.F Struc.Biol.Crystalliz.Commun.63:1029,2007
OSTI ID:954007
Past work has shown that it is feasible to mutate surface residues of soluble proteins and to a lesser extent membrane proteins in order to improve their crystallization behavior. Described here is a successful application of this approach to the integral membrane protein Thermus thermophilus cytochrome ba(3) oxidase. Two mutant forms of this enzyme (I-K258R and I-K258R/II-E4Q) were created in which symmetrical crystal contacts within crystals of wild-type enzyme were modified. These mutant proteins had greatly shortened crystallization times, decreasing from approximately 30 d for the wild type to 1-3 d for the mutants, and crystallization was highly reproducible. Native-like proteins crystallize in space group P4(3)2(1)2, whereas the mutant proteins crystallize in space group P4(1)2(1)2 with a different packing arrangement. Crystals of the P4(3)2(1)2 form occasionally diffracted to 2.4-2.3 A resolution following controlled dehydration, while those of the P4(1)2(1)2 form routinely diffracted to between 3.0 and 2.6 A for crystals that had been cryoprotected but not dehydrated.
- Research Organization:
- Stanford Linear Accelerator Center (SLAC)
- Sponsoring Organization:
- USDOE
- DOE Contract Number:
- AC02-76SF00515
- OSTI ID:
- 954007
- Report Number(s):
- SLAC-REPRINT-2009-385
- Journal Information:
- Acta Crystallogr.F Struc.Biol.Crystalliz.Commun.63:1029,2007, Journal Name: Acta Crystallogr.F Struc.Biol.Crystalliz.Commun.63:1029,2007 Journal Issue: 12 Vol. 63; ISSN 1744-3091
- Country of Publication:
- United States
- Language:
- English
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