Solubilization and partial characterization of phospholipase A from rat heart sarcoplasmic reticulum.
Phospholipase A has been solubilized from the sarcoplasmic reticulum of rat heart by treatment with Tris buffer, potassium chloride, taurodeoxycholate or octyl glucoside. On HPLC gel permeation, two phospholipases were identified at the void volume of a TSK 3000 column and at an apparent molecular mass of 60 kDa. The two activity peaks exhibited a predominance of phospholipase A1 activity (83-91%) and a lesser phospholipase C activity (4-9%) using sonicated 1-palmitoyl-2[1-14C]oleoylphosphatidylcholine liposomes as substrate. The voiding phospholipase A peak, which represented the bulk of the recovered activity, exhibited a requirement for calcium ions in the 0.3-3 microM range. The heat stability and response to mercuric ions was studied and some similarities were noted between the solubilized sarcoplasmic reticulum phospholipases A and the cytosolic phospholipases A of rat heart. It is speculated that the cytosolic phospholipase A which we reported earlier may represent in part phospholipase A released from sarcoplasmic reticulum during isolation of the subcellular membrane fractions.
Trotz
M
M
Department of Medicine, University of California, San Diego.
Hein
L
L
Hostetler
K Y
KY
D000818:Animals; D002118:Calcium; D003600:Cytosol / Q000201:enzymology; D008297:Male; D009206:Myocardium / Q000201:enzymology*; D010740:Phospholipases / Q000302:isolation & purification*; D010741:Phospholipases A / Q000302:isolation & purification* / Q000378:metabolism; D054466:Phospholipases A1; D051381:Rats; D011919:Rats, Inbred Strains; D012519:Sarcoplasmic Reticulum / Q000201:enzymology*; D012995:Solubility; D013379:Substrate Specificity; D010738:Type C Phospholipases / Q000378:metabolism
D010740:Phospholipases; D010741:Phospholipases A; D054466:Phospholipases A1; D010738:Type C Phospholipases; D002118:Calcium