Isolation, characterization and localization of annexin V from chicken liver.
Annexin V has been purified from chicken liver; 40 mg of annexin V was obtained per kg of tissue. In contrast with mammalian liver, very little annexin VI was obtained. Surprisingly, chicken liver annexin V resembles mammalian annexin IV in its M(r) (32,500) and its isoelectric point (5.6), but amino-acid-sequence analysis demonstrates identity with chicken annexin V (anchorin CII). It binds to phospholipids in a Ca(2+)-dependent manner with free-Ca2+ concentrations for half-maximal binding to phosphatidylserine and phosphatidic acid of 10 microM; phosphatidylethanolamine of 32 microM and phosphatidylinositol of 90 microM. No binding to phosphatidylcholine was observed at Ca2+ concentrations up to 300 microM. In isolated liver membranes a significant proportion of annexin V was not extractable with EGTA but could only be extracted with Triton X-100, suggesting the existence of a tightly membrane-associated form of annexin V. A specific antiserum to chicken annexin V was used to localize the protein in adult and embryonic chicken liver. In the adult, annexin V was highly concentrated in epithelial cells lining the bile ducts, and along the bile canaliculi. In embryonic liver, strong staining of the bile-duct epithelial cells was again evident, and in addition, endothelial cells were strongly immunoreactive.
Boustead
C M
CM
Department of Biochemistry and Molecular Biology, University of Leeds, U.K.
Brown
R
R
Walker
J H
JH
D000595:Amino Acid Sequence; D000818:Animals; D017304:Annexin A5 / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D002118:Calcium / Q000494:pharmacology; D002645:Chickens; D002848:Chromatography, DEAE-Cellulose; D004591:Electrophoresis, Polyacrylamide Gel; D005895:Gizzard, Avian / Q000737:chemistry; D007124:Immunoenzyme Techniques; D008099:Liver / Q000737:chemistry*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010743:Phospholipids / Q000378:metabolism; D014018:Tissue Distribution
D017304:Annexin A5; D010743:Phospholipids; D002118:Calcium