Crystallization and preliminary X-ray diffraction studies of human procathepsin L.
Human procathepsin L has been expressed in the yeast Pichia pastoris and its inactive (Cys25Ser) and unglycosylated (Thr110Ala) mutant purified, concentrated to 4 mg/ml, and crystallized by vapor diffusion against solution containing 1.4 M (Na,K)PO4 buffer, pH 7.8. Crystal size was increased by multiple macroseeding. The crystals are orthorhombic, of space group P2 1 2 1 2 1, with cell dimensions of a = 40.2 A, b = 88.4 A, and c = 94.9 A. A 2.2 A native data set was collected using synchrotron radiation. Although molecular replacement solution for the mature portion of the enzyme was easily found, the resulting maps could not be interpreted in the proregion. Heavy-atom derivative search is in progress.
Coulombe
R
R
Protein Engineering Network of Centers of Excellence, Canada.
Li
Y
Y
Takebe
S
S
Ménard
R
R
Mason
P
P
Mort
J S
JS
Cygler
M
M
D056668:Cathepsin L; D002403:Cathepsins / Q000037:antagonists & inhibitors / Q000737:chemistry*; D018360:Crystallography, X-Ray; D018076:DNA, Complementary; D004058:Diffusion; D004792:Enzyme Precursors / Q000037:antagonists & inhibitors / Q000737:chemistry*; D006801:Humans; D011994:Recombinant Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry
D018076:DNA, Complementary; D004792:Enzyme Precursors; D011994:Recombinant Proteins; D002403:Cathepsins; C063905:procathepsin L; D056668:Cathepsin L