5'-Nucleotidases of chicken gizzard and human pancreatic adenocarcinoma cells are anchored to the plasma membrane via a phosphatidylinositol-glycan.
We have analysed the membrane anchorage of plasma-membrane 5'-nucleotidase, an ectoenzyme which can mediate binding to components of the extracellular matrix. We demonstrated that the purified enzyme obtained from chicken gizzard and a human pancreatic adenocarcinoma cell line were both completely transformed into a hydrophilic form by treatment with phospholipases C and D, cleaving glycosylphosphatidylinositol (GPI). These data indicate the presence of a glycolipid linker employed for membrane anchoring of the 5'-nucleotidase obtained from both sources. Incubation of plasma membranes under identical conditions revealed that about half of the AMPase activity was resistant to GPI-hydrolysing phospholipases. Investigation of the enzymic properties of purified chicken gizzard 5'-nucleotidase revealed only minor changes after removal of the phosphatidylinositol linker. However, cleavage of the membrane anchor resulted in an increased sensitivity towards inhibition by concanavalin A. After tissue fractionation, chicken gizzard 5'-nucleotidase could be obtained as either a membrane-bound or a soluble protein; the latter is suspected to be released from the plasma membrane by endogenous phospholipases. Higher-molecular-mass proteins immuno-cross-reactive with the purified chicken gizzard 5'-nucleotidase were detected as both soluble and membrane-bound forms.
Stochaj
U
U
Institut für Anatomie und Zellbiologie, Marburg, Federal Republic of Germany.
Flocke
K
K
Mathes
W
W
Mannherz
H G
HG
D015720:5'-Nucleotidase / Q000378:metabolism*; D000230:Adenocarcinoma / Q000201:enzymology*; D000818:Animals; D002462:Cell Membrane / Q000378:metabolism; D002645:Chickens; D005895:Gizzard, Avian; D006801:Humans; D009130:Muscle, Smooth / Q000201:enzymology*; D009708:Nucleotidases / Q000378:metabolism; D010190:Pancreatic Neoplasms / Q000201:enzymology*; D010716:Phosphatidylinositols / Q000378:metabolism; D011134:Polysaccharides / Q000378:metabolism; D011485:Protein Binding
D010716:Phosphatidylinositols; D011134:Polysaccharides; D009708:Nucleotidases; C018775:adenosine monophosphatase; D015720:5'-Nucleotidase