CRYSTAL STRUCTURE OF a DUF2118 family protein (MMP0046) FROM METHANOCOCCUS MARIPALUDIS AT 2.20 A RESOLUTION

Joint Center for Structural Genomics (JCSG)

Crystal structure of Domain of Unknown Function from the Pfam-B_34464 Family (NP_987166.1) from Methanococcus maripaludis JJ (DSM 2067) at 2.20 A resolution

FLAT COLLIMATING MIRROR, TOROID FOCUSING MIRROR

Domain of Unknown Function from the Pfam-B_34464 Family

(MSE)GSDKIHHHHHHENLYFQG(MSE)KIPKIYVEGELNDGDRVAIEKDGNAIIFLEKDEEYSGNGKLLYQVIYDDLAK
Y(MSE)SLDTLKKDVLIQYPDKHTLTYLKAGTKLISVPAEGYKVYPI(MSE)DFGFRVLKGYRLATLESKKGDLRYVNSP
VSGTVIF(MSE)NEIPSERANYVFY(MSE)LEE

MGSDKIHHHHHHENLYFQGMKIPKIYVEGELNDGDRVAIEKDGNAIIFLEKDEEYSGNGKLLYQVIYDDLAKYMSLDTLK
KDVLIQYPDKHTLTYLKAGTKLISVPAEGYKVYPIMDFGFRVLKGYRLATLESKKGDLRYVNSPVSGTVIFMNEIPSERA
NYVFYMLEE

21.5% polyethylene glycol 3350, 0.214M ammonium citrate, NANODROP, VAPOR DIFFUSION, SITTING DROP, temperature 277K

THE CONSTRUCT WAS EXPRESSED WITH A PURIFICATION TAG MGSDKIHHHHHHENLYFQG. THE STRAIN CLONED, METHANOCOCCUS MARIPALUDIS JJ, DIFFERS FROM THE DATABASE SEQUENCE STRAIN, METHANOCOCCUS MARIPALUDIS S2. SEQUENCING OF THE CLONED CONSTRUCT SHOWS A LEUCINE AT POSITION 74 INSTEAD OF A PHENYLALANINE. THE LEUCINE AT POSITION 74 IS SUPPORTED BY THE ELECTRON DENSITY.

1. HYDROGENS HAVE BEEN ADDED IN THE RIDING POSITIONS. 2. ATOM RECORD CONTAINS RESIDUAL B FACTORS ONLY. 3. 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. 3. EDO WAS MODELED BASED ON CRYO CONDITIONS. 4. RAMACHANDRAN OUTLIERS A50 AND C50 ARE IN GOOD DENSITY.

AUTHORS STATE THAT THE PROTOMER MAY FORM A DIMER BASED ON CRYSTAL PACKING ANALYSIS. ANALYTICAL SIZE EXCLUSION CHROMATOGRAPHY SUPPORTS THE ASSIGNMENT OF A DIMER AS A SIGNIFICANT OLIGOMERIC STATE.

STRUCTURAL GENOMICS, JOINT CENTER FOR STRUCTURAL GENOMICS, JCSG, PROTEIN STRUCTURE INITIATIVE, PSI-2, UNKNOWN FUNCTION

MKIPKIYVEGELNDGDRVAIEKDGNAIIFLEKDEEYSGNGKLLYQVIYDDLAKYMSLDTLKKDVLIQYPDKHTFTYLKAG
TKLISVPAEGYKVYPIMDFGFRVLKGYRLATLESKKGDLRYVNSPVSGTVIFMNEIPSERANYVFYMLEE