Crystallization and initial crystallographic results for pepstatin A inhibited bovine cathepsin D.
Cathepsin D was purified from bovine liver by a method using two pepstatin A affinity columns. The eluted protein was combined with pepstatin A and the complex crystallized from 15% polyethylene glycol 8000 at pH 5.9. The crystals diffract to a resolution of 3.0 A and have space group P2(1)2(1)2(1) with unit cell dimensions a = 74.8 A, b = 76.0 A, c = 157.7 A. There are two molecules in the asymmetric unit. The structure was solved by molecular replacement using a pepsin search model and both molecules showed clearly interpretable density in the position expected for pepstatin A in a preliminary difference map. The refined model has r.m.s. deviations from ideal bond lengths and angles of 0.014 A and 3.2 degrees, respectively, and a crystallographic R factor of 17%.
Bieber
F
F
European Molecular Biology Laboratory, Heidelberg, Germany.
Brachvogel
V
V
Arni
R
R
Fusek
M
M
Metcalf
P
P
D000818:Animals; D002402:Cathepsin D / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000302:isolation & purification; D002417:Cattle; D003460:Crystallization; D008099:Liver / Q000737:chemistry; D010436:Pepstatins / Q000737:chemistry / Q000494:pharmacology*; D014961:X-Ray Diffraction
D010436:Pepstatins; C014195:Streptomyces pepsin inhibitor; D002402:Cathepsin D; C031375:pepstatin