Both tissue and serum phospholipases release rat intestinal alkaline phosphatase.
Rat intestinal alkaline phosphatase (IAP) is unique among the brush-border membrane enzymes in that it is released bidirectionally (lumen and blood) and exists in either soluble (serum) or particulate (cellular) form. To elucidate the mechanism of membrane release, we examined the effects of phosphatidylinositol-specific phospholipase C (PtdIns-PLC) and serum anchor-specific phospholipase D (PLD) on the solubility of the various tissue forms of IAP. The "solubility" of cytosol IAP could be explained in part by intracellular PtdIns-PLC activity, detected by production of acidic IAP isomers, and by ethylene glycol-bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid (EGTA)-sensitive PtdIns hydrolysis. Contamination with serum (abundant with anchor-specific PLD) was responsible for the complete or partial solubilization of IAP that was found during processing of light mucosal scrapings. Anchor-specific PLD activity was increased after fat feeding, and the IAP released did not react with antiserum that recognizes the PtdIns-PLC-released phospholipid portion of trypanosomal variable surface glycoprotein. These data are consistent with the hypothesis that, after secretion from the enterocyte bound to a phospholipid-rich membranous particle, IAP release into serum is mediated by serum anchor-specific PLD. The soluble forms of IAP in the lumen and the cytosol fraction appear to be due to a combination of endogenous PtdIns-PLC activity and anchor-specific PLD contamination that occurs during cell fractionation.
Eliakim
R
R
Department of Medicine, Washington University School of Medicine, St. Louis, Missouri 63110.
Becich
M J
MJ
Green
K
K
Alpers
D H
DH
D000469:Alkaline Phosphatase / Q000378:metabolism*; D000818:Animals; D007413:Intestinal Mucosa / Q000201:enzymology* / Q000648:ultrastructure; D007421:Intestine, Small / Q000201:enzymology; D007527:Isoenzymes / Q000097:blood / Q000378:metabolism*; D007700:Kinetics; D008297:Male; D016253:Microscopy, Immunoelectron; D008871:Microvilli / Q000201:enzymology* / Q000648:ultrastructure; D010740:Phospholipases / Q000097:blood / Q000378:metabolism*; D051381:Rats; D011919:Rats, Inbred Strains
D007527:Isoenzymes; D010740:Phospholipases; D000469:Alkaline Phosphatase