Protein ID (e.g. MH) PP-BRD19
SGC Clone ID construct ID from Polymorph followed by plate ID and plate location) LdBPK_363520.1:MAC055-E04:F253289
NCBI accession (or equivalent)
Vector pET15-MHL
Construct comments (e.g. mutations, etc.) his tag cut
Construct sequence HPRPLPAGKHAHRQSLETIPEVAELYHCIYKLYNEEESSVWFREPVNALAQEIFTYYDVVKSPMSLRHILDNIVKGDTYSTALQVMEDVELIWKNCITFNGANSLLATEAGKCRSALDRIRRAYQDDQRITVEEAE
Expression Host (e.g. BL21 (DE3)) BL21-CodonPlus(DE3)-RIL
Purification buffers Binding Buffer: 50 mM HEPES pH 7.5, 500 mM NaCl, 5 mM imidazole, and 5 % glycerol
Wash Buffer: 50 mM HEPES pH 7.5, 500 mM NaCl, 30 mM imidazole, and 5 % glycerol
Elution Buffer: 50 mM HEPES pH 7.5, 500 mM NaCl, 250 mM imidazole, and 5 % glycerol
Gel Filtration buffer: 20 mM HEPES, pH 7.5, 150 mM NaCl
Crystallization plate, well, drop ( e.g. MAY024:A2-1 for plate MAY024, well A2, drop 1) IC-MAZ7PF, E8-1
Cryo conditions 10% ethylene glycol
Crystallization procedure (if above conditions are not sufficient) protein concentration:15mg/ml in 20mM HEPES7.5 and 150 mM NaCl.
Crystallization The protein was crystallized at 20 ºC in 30%PEG2000 MME, 0.15M KBr with bromosporine using the Sitting drop method.
