Purification, crystallization, and preliminary X-ray diffraction analysis of rat kidney annexin V, a calcium-dependent phospholipid-binding protein.
We have purified annexin V, a monomeric 35-kDa protein, from rat kidney using calcium-dependent phospholipid chromatography. The identity of annexin V was confirmed by immunoblot analysis using monospecific anti-annexin V antibody. Large single crystals of annexin V in the presence of calcium have been grown from ammonium sulfate under a variety of conditions, with an optimum pH range of 7.5-8.0. The crystals diffract to at least 2.2 A Bragg spacing and are stable to x-rays. Preliminary crystallographic analysis reveals the space group to be R3, with hexagonal cell dimensions of a = b = 156.8 A and c = 36.9 A, and there is one molecule/asymmetric unit.
Seaton
B A
BA
Department of Physiology, Boston University School of Medicine, Massachusetts 02118.
Head
J F
JF
Kaetzel
M A
MA
Dedman
J R
JR
D000818:Animals; D002118:Calcium / Q000494:pharmacology*; D002135:Calcium-Binding Proteins / Q000302:isolation & purification*; D002846:Chromatography, Affinity; D003460:Crystallization; D006863:Hydrogen-Ion Concentration; D007668:Kidney / Q000032:analysis*; D010743:Phospholipids / Q000378:metabolism*; D051381:Rats; D014961:X-Ray Diffraction
D002135:Calcium-Binding Proteins; D010743:Phospholipids; D002118:Calcium