Identification of novel binding partners (annexins) for the cell death signal phosphatidylserine and definition of their recognition motif.
Identification and clearance of apoptotic cells prevents the release of harmful cell contents thereby suppressing inflammation and autoimmune reactions. Highly conserved annexins may modulate the phagocytic cell removal by acting as bridging molecules to phosphatidylserine, a characteristic phagocytosis signal of dying cells. In this study five members of the structurally and functionally related annexin family were characterized for their capacity to interact with phosphatidylserine and dying cells. The results showed that AnxA3, AnxA4, AnxA13, and the already described interaction partner AnxA5 can bind to phosphatidylserine and apoptotic cells, whereas AnxA8 lacks this ability. Sequence alignment experiments located the essential amino residues for the recognition of surface exposed phosphatidylserine within the calcium binding motifs common to all annexins. These amino acid residues were missing in the evolutionary young AnxA8 and when they were reintroduced by site directed mutagenesis AnxA8 gains the capability to interact with phosphatidylserine containing liposomes and apoptotic cells. By defining the evolutionary conserved amino acid residues mediating phosphatidylserine binding of annexins we show that the recognition of dying cells represent a common feature of most annexins. Hence, the individual annexin repertoire bound to the cell surface of dying cells may fulfil opsonin-like function in cell death recognition.
Medical Faculty, Center for Biochemistry, University of Cologne, 50931 Cologne, Germany.
Sabrina
S
Rosenbaum
Sandra
S
Kreft
Julia
J
Etich
Christian
C
Frie
Jacek
J
Stermann
Ivan
I
Grskovic
Benjamin
B
Frey
Dirk
D
Mielenz
Ernst
E
Pöschl
Udo
U
Gaipl
Mats
M
Paulsson
Bent
B
Brachvogel
D020816:Amino Acid Motifs; D000818:Animals; D017302:Annexins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D017209:Apoptosis / Q000502:physiology*; D002118:Calcium / Q000378:metabolism*; D019143:Evolution, Molecular; D051379:Mice; D041681:NIH 3T3 Cells; D010718:Phosphatidylserines / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding; D015398:Signal Transduction / Q000502:physiology*
D017302:Annexins; D010718:Phosphatidylserines; D002118:Calcium