Crystallization and preliminary X-ray analysis of the atrial natriuretic peptide (ANP) receptor extracellular domain complex with ANP: use of ammonium sulfate as the cryosalt.
Atrial natriuretic peptide (ANP) plays a major role in blood pressure and volume regulation owing to its natriuretic and vasodilatory activities. The ANP receptor is a single-span transmembrane receptor coupled to its intrinsic guanylyl cyclase activity. The extracellular hormone-binding domain of rat ANP receptor (ANPR) was overexpressed by permanent transfection in CHO cells and purified. ANPR complexed with ANP was crystallized at 301 K by the hanging-drop vapor-diffusion method. The crystals were frozen in 3.4 M ammonium sulfate used as a cryoprotectant. The crystals diffracted to 3.1 A resolution using synchrotron radiation and belonged to the hexagonal space group P6(1), with unit-cell parameters a = b = 100.3, c = 258.6 A.
D000645:Ammonium Sulfate / Q000737:chemistry*; D000818:Animals; D009320:Atrial Natriuretic Factor / Q000737:chemistry*; D016466:CHO Cells; D006224:Cricetinae; D003460:Crystallization / Q000379:methods; D018360:Crystallography, X-Ray; D005110:Extracellular Space / Q000737:chemistry; D008958:Models, Molecular; D017434:Protein Structure, Tertiary; D051381:Rats; D017461:Receptors, Atrial Natriuretic Factor / Q000096:biosynthesis / Q000737:chemistry*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry; D055672:Static Electricity
D011994:Recombinant Proteins; D009320:Atrial Natriuretic Factor; D017461:Receptors, Atrial Natriuretic Factor; D000645:Ammonium Sulfate