Preliminary X-ray Diffraction Analysis of the Cytoplasmic N-terminal Domain of the Na/HCO3 Cotransporter NBCe1-A
The N-terminal cytoplasmic domain of the Na{sup +}-coupled HCO{sub 3}{sup -} cotransporter NBCe1-A (NtNBCe1) has been linked with proximal renal tubular acidosis. In a previous purification study of recombinant NtNBCe1, crystal growth at a suboptimal protein concentration (<1 mg ml{sup -1}) yielded small single diamond-shaped crystals that diffracted poorly. In the present study, by increasing the protein concentration 50-fold, the crystal size was doubled and robustness was also improved. Crystal annealing made the crystals suitable for X-ray diffraction. The crystals either belong to space group P3121 or P31 with pseudo P3121 symmetry, with unit-cell parameters a = 51.7, b = 51.7, c = 200.6 Angstroms, {alpha} = {beta} = 90, {gamma} = 120 deg, and diffract X-rays to 3.0 Angstroms resolution. The calculated Matthews number is 1.9 Angstroms{sup 3} Da{sup -1}, with two monomers of molecular weight {approx}83 kDa in the asymmetric unit. The molecular- replacement packing solution shows that the molecules form dimers by a domain-swapping mechanism.
- Research Organization:
- Brookhaven National Lab. (BNL), Upton, NY (United States). National Synchrotron Light Source
- Sponsoring Organization:
- Doe - Office Of Science
- DOE Contract Number:
- DE-AC02-98CH10886
- OSTI ID:
- 914352
- Report Number(s):
- BNL-78920-2007-JA; TRN: US200809%%194
- Journal Information:
- Acta Cryst. F, Vol. 62
- Country of Publication:
- United States
- Language:
- English
Similar Records
Expression and Purification of the Cytoplasmic N-Terminal Domain of the Na/HCO3 Cotransporter NBCe1-A: Structural Insights from the a Generalized Approach
Crystallization and X-ray data analysis of the 10 kDa C-terminal lid subdomain from Caenorhabditis elegans Hsp70