Effect of organic anions on the crystallization of the Ca2(+)-ATPase of muscle sarcoplasmic reticulum.
The effect of varying the solute species on the crystallization of the Ca2(+)-ATPase from rabbit muscle reticulum (SR) is reported. We have found that substitution of KCl with salts of organic acids in the crystallization protocol reported by Pikula et al. has a profound effect on the size of two-dimensional crystalline arrays. Crystalline arrays of up to 3 microns diameter have been obtained by incubating purified calcium ATPase in standard crystallization medium but with 0.8 M sodium propionate substituted for KCl. These two-dimensional (2-D) arrays display a reduced tendency to stack in addition to having larger planar dimensions. Increasing the KCl concentration does not have the same effect on stacking or crystal growth that sodium propionate has. The production of 2-D sheets has some dependence on the hydrocarbon chain length of the salt because crystals formed in propionate were larger and less stacked than those formed in acetate or formate. There seems to be no dependence on cation. These observations suggest that in addition to reducing the forces that lead to stacking of the sheets, propionate may facilitate incorporation of the detergent-solubilized protein into the 2-D sheet.
Misra
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M
Department of Cell Biology, Duke University Medical Center, Durham, NC 27710.
Taylor
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Oliver
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Taylor
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D000085:Acetates / Q000494:pharmacology; D000818:Animals; D000838:Anions; D000252:Calcium-Transporting ATPases / Q000737:chemistry* / Q000302:isolation & purification / Q000648:ultrastructure; D002264:Carboxylic Acids / Q000494:pharmacology*; D003460:Crystallization; D008854:Microscopy, Electron; D009132:Muscles / Q000201:enzymology; D009994:Osmolar Concentration; D011189:Potassium Chloride / Q000494:pharmacology; D011817:Rabbits; D012519:Sarcoplasmic Reticulum / Q000201:enzymology* / Q000648:ultrastructure
D000085:Acetates; D000838:Anions; D002264:Carboxylic Acids; D011189:Potassium Chloride; D000252:Calcium-Transporting ATPases; C018824:ammonium acetate