A series of annexins from human placenta and their characterization by use of an endogenous phospholipase A2.
Membranes from human placenta contain proteins which inhibit the activity of phospholipases A2 by binding to phospholipid thus impeding substrate availability. We used unilamellar mixed liposomes and a partially purified cytosolic phospholipase A2 from placenta for characterizing this substrate-depleting activity. A major portion of these inhibitory proteins was released by extracting washed membranes with a Ca+(+)-chelator. Biochemical fractionation and systematic analysis resulted in the unequivocal identification of a series of annexin proteins. We describe a straightforward procedure which allows to obtain 8 annexins from placenta either in pure form or as a mixture of two annexins. One of them was obtained in two forms which had the same molecular mass of 68 kDa but differed in charge. We also present suggestive evidence for a novel annexin I-related polypeptide of Mr 45,000 which is an excellent in vitro substrate for protein kinase C. We estimate that about 2% of the total placental membrane proteins are annexins. For achieving half inhibition of phospholipase A2 activity with pure annexins, up to a 6.5-fold difference in the amounts of protein was observed when calculated on a molar basis. This suggests specificity of individual annexin species.
Buhl
W J
WJ
Institut für Biologische Chemie Universität, Heidelberg/Bundesrepublik Deutschland.
García
M T
MT
Zipfel
M
M
Schiebler
W
W
Gehring
U
U
D017302:Annexins; D002135:Calcium-Binding Proteins / Q000737:chemistry* / Q000145:classification / Q000302:isolation & purification / Q000494:pharmacology / Q000502:physiology; D005260:Female; D006023:Glycoproteins / Q000737:chemistry* / Q000145:classification / Q000502:physiology; D006801:Humans; D008081:Liposomes / Q000378:metabolism; D008565:Membrane Proteins / Q000737:chemistry* / Q000145:classification / Q000502:physiology; D008566:Membranes / Q000737:chemistry / Q000502:physiology; D010741:Phospholipases A / Q000037:antagonists & inhibitors* / Q000187:drug effects; D054467:Phospholipases A2; D010920:Placenta / Q000737:chemistry* / Q000502:physiology; D011247:Pregnancy
D017302:Annexins; D002135:Calcium-Binding Proteins; D006023:Glycoproteins; D008081:Liposomes; D008565:Membrane Proteins; D010741:Phospholipases A; D054467:Phospholipases A2