Annexins I, II and III are specific choline binding proteins.
We have isolated choline binding proteins from the plasma membrane fraction fraction of human lung epithelium-derived cell line (A549) by means of detergent solubilization, anion exchange and affinity chromatography. One of the affinity purified proteins had a specific choline binding activity of 44-57 pmol/mg, representing a two to three hundredfold enrichment relative to the specific activity of freshly prepared plasma membranes. The purified protein has a molecular mass of 38 kDa by SDS PAGE analysis and was identified as annexin II by N-terminal microsequencing. Annexin II, however, had not previously been known for choline binding activity. We therefore prepared a mixture of authentic annexins (I-V) from A549 cells. The mixture had a choline binding activity of 15 to 18 pmol/mg. The annexin mixture was subsequently affinity chromatographed on the choline-conjugated Sepharose 6B column. Analyses by SDS PAGE and immunoblot revealed that annexins I, II, and III are bound to the choline column while annexins IV and V did not. These results indicate that some of the annexins have specific choline binding activities.
Zimmerman
U J
UJ
Institute for Environmental Medicine, University of Pennsylvania School of Medicine, Philadelphia 19104, USA.
Hennigan
B B
BB
Liu
L
L
Campbell
C H
CH
Fisher
A B
AB
D000595:Amino Acid Sequence; D000818:Animals; D017305:Annexin A1 / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D017306:Annexin A2 / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D017318:Annexin A3 / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D002154:Calpain / Q000302:isolation & purification / Q000378:metabolism; D002417:Cattle; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism*; D002794:Choline / Q000378:metabolism*; D002846:Chromatography, Affinity; D002852:Chromatography, Ion Exchange; D004848:Epithelium; D004912:Erythrocytes / Q000201:enzymology; D006801:Humans; D008168:Lung; D008969:Molecular Sequence Data; D008970:Molecular Weight
D017305:Annexin A1; D017306:Annexin A2; D017318:Annexin A3; D002154:Calpain; D002794:Choline