Optimization of Met8p crystals through protein-storage buffer manipulation.
Sirohaem, the prosthetic group of assimilatory sulfite and nitrite reductases, is a modified tetrapyrrole that belongs to the same fraternity of metallo-prosthetic groups as haem, chlorophyll, cobalamin and coenzyme F430 [Warren & Scott (1990), Trends Biochem Sci. 15, 486-491]. In Saccharomyces cerevisiae, the last step in the biosynthesis of sirohaem involves Met8p, a bifunctional enzyme responsible for both the NAD(+)-dependent dehydrogenation of the corrin ring and ferrochelation. Optimization of the protein storage buffer according to the results of crystallization trials resulted in a more monodisperse protein solution. Crystals were grown that diffracted to 2.1 A.
Schubert
H L
HL
Department of Biochemistry, University of Utah, Salt Lake City, UT 84132, USA. heidi@snowbird.med.utah.edu
Raux
E
E
Warren
M J
MJ
Wilson
K S
KS
D002021:Buffers; D003460:Crystallization; D018360:Crystallography, X-Ray; D005294:Ferrochelatase; D005656:Fungal Proteins / Q000737:chemistry*; D009097:Multienzyme Complexes / Q000737:chemistry; D011487:Protein Conformation; D012441:Saccharomyces cerevisiae / Q000737:chemistry / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins
D002021:Buffers; D005656:Fungal Proteins; D009097:Multienzyme Complexes; D029701:Saccharomyces cerevisiae Proteins; C103747:MET8 protein, S cerevisiae; D005294:Ferrochelatase