Crystal structure of a Putative ubiquinone biosynthesis protein (Npun02000094) from Nostoc punctiforme PCC 73102 at 2.85 A resolution

Joint Center for Structural Genomics (JCSG)

Single crystal Si(111) bent monochromator (horizontal focusing)

Putative ubiquinone biosynthesis protein

G(MSE)ENVITYNNDKGLLAYIQFLASSAQKIGDGNTDRVFDFEDALDQTQ(MSE)AQLAVDELKKIPEVNALFSERWLP
APFNLDDLAKLPEGTLGHVYARE(MSE)KARGFDPYFYKKVPVVDDISYLK(MSE)LWRSTHDIYHVVAGFDTNVFGEIG
LQAFFLAQTPIPISV(MSE)LLSFG(MSE)V(MSE)ISLYQPTNFKAL(MSE)TEISRGYRVGSHTPGKLIAQKWDQLWD
VQVSEIRERLGVNSIL

GMENVITYNNDKGLLAYIQFLASSAQKIGDGNTDRVFDFEDALDQTQMAQLAVDELKKIPEVNALFSERWLPAPFNLDDL
AKLPEGTLGHVYAREMKARGFDPYFYKKVPVVDDISYLKMLWRSTHDIYHVVAGFDTNVFGEIGLQAFFLAQTPIPISVM
LLSFGMVMISLYQPTNFKALMTEISRGYRVGSHTPGKLIAQKWDQLWDVQVSEIRERLGVNSIL

DATA WERE SCALED USING XSCALE WITH FRIEDEL PAIRS KEPT AS SEPARATE WHEN COMPUTING R-SYM, COMPLETENESS AND <I/SIGMA(I)>

20.0000% polyethylene glycol 3000, 0.1M citric acid pH 5.5, Additive: 0.001 M Ubiquinone, NANODROP', VAPOR DIFFUSION, SITTING DROP, temperature 293K

THE CONSTRUCT WAS EXPRESSED WITH A PURIFICATION TAG MGSDKIHHHHHHENLYFQG. THE TAG WAS REMOVED WITH TEV PROTEASE LEAVING ONLY A GLYCINE (0) FOLLOWED BY THE TARGET SEQUENCE.

1. A MET-INHIBITION PROTOCOL WAS USED FOR SELENOMETHIONINE INCORPORATION DURING PROTEIN EXPRESSION. THE OCCUPANCY OF THE SE ATOMS IN THE MSE RESIDUES WAS REDUCED TO 0.75 FOR THE REDUCED SCATTERING POWER DUE TO PARTIAL S-MET INCORPORATION. 2. B-FACTORS CONTAIN BOTH TLS AND RESIDUAL COMPONENTS.

Coq4 superfamily, Structural Genomics, Joint Center for Structural Genomics, JCSG, Protein Structure Initiative, PSI-2, BIOSYNTHETIC PROTEIN

MENVITYNNDKGLLAYIQFLASSAQKIGDGNTDRVFDFEDALDQTQMAQLAVDELKKIPEVNALFSERWLPAPFNLDDLA
KLPEGTLGHVYAREMKARGFDPYFYKKVPVVDDISYLKMLWRSTHDIYHVVAGFDTNVFGEIGLQAFFLAQTPIPISVML
LSFGMVMISLYQPTNFKALMTEISRGYRVGSHTPGKLIAQKWDQLWDVQVSEIRERLGVNSIL