The copines, a novel class of C2 domain-containing, calcium-dependent, phospholipid-binding proteins conserved from Paramecium to humans.
In an attempt to identify proteins that might underlie membrane trafficking processes in ciliates, calcium-dependent, phospholipid-binding proteins were isolated from extracts of Paramecium tetraurelia. The major protein obtained, named copine, had a mass of 55 kDa, bound phosphatidylserine but not phosphatidylcholine at micromolar levels of calcium but not magnesium, and promoted lipid vesicle aggregation. The sequence of a 920-base pair partial cDNA revealed that copine is a novel protein that contains a C2 domain likely to be responsible for its membrane active properties. Paramecium was found to have two closely related copine genes, CPN1 and CPN2. Current sequence data bases indicate the presence of multiple copine homologs in green plants, nematodes, and humans. The full-length sequences reveal that copines consist of two C2 domains at the N terminus followed by a domain similar to the A domain that mediates interactions between integrins and extracellular ligands. A human homolog, copine I, was expressed in bacteria as a fusion protein with glutathione S-transferase. This recombinant protein exhibited calcium-dependent phospholipid binding properties similar to those of Paramecium copine. An antiserum raised against a fragment of human copine I was used to identify chromobindin 17, a secretory vesicle-binding protein, as a copine. This association with secretory vesicles, as well the general ability of copines to bind phospholipid bilayers in a calcium-dependent manner, suggests that these proteins may function in membrane trafficking.
Creutz
C E
CE
Department of Pharmacology, University of Virginia, Charlottesville 22908, USA. Creutz@virginia.edu
Tomsig
J L
JL
Snyder
S L
SL
Gautier
M C
MC
Skouri
F
F
Beisson
J
J
Cohen
J
J
D000595:Amino Acid Sequence; D000818:Animals; D017302:Annexins; D001483:Base Sequence; D002118:Calcium / Q000378:metabolism; D002352:Carrier Proteins / Q000737:chemistry* / Q000378:metabolism; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000737:chemistry; D016054:DNA, Protozoan / Q000737:chemistry; D015723:Gene Library; D006801:Humans; D008565:Membrane Proteins / Q000378:metabolism; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009348:Nematoda; D016805:Paramecium tetraurelia; D010743:Phospholipids / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry; D016415:Sequence Alignment
D017302:Annexins; D002352:Carrier Proteins; D018076:DNA, Complementary; D016054:DNA, Protozoan; D008565:Membrane Proteins; D010743:Phospholipids; D011994:Recombinant Proteins; C110203:copine; D002118:Calcium