Cloning and expression of cDNA for human endonexin II, a Ca2+ and phospholipid binding protein.
Endonexin II is a member of the family of Ca2+-dependent phospholipid binding proteins known as annexins. We cloned human endonexin II cDNA and expressed it in Escherichia coli. The apparent size and Ca2+-dependent phospholipid binding properties of purified recombinant endonexin II were indistinguishable from those of the placental protein. A single mRNA of approximately 1.6 kilobase pairs was found to be expressed in human cell lines and placenta and was in close agreement with the length of the cDNA clone (1.59 kilobase pairs). The cDNA predicted a 320-amino acid protein with a sequence that was in agreement with the previously determined partial amino acid sequence of endonexin II isolated from placenta. Endonexin II contained 58, 46, and 43% sequence identity to protein II, calpactin I (p36, protein I), and lipocortin I (p35), respectively. The partial sequence of bovine endonexin I was aligned with the sequence of endonexin II to give 63% sequence identity. Like these other proteins, endonexin II had a 4-fold internal repeat of approximately 70 residues preceded by an amino-terminal domain lacking similarity to the repeated region. It also had significant sequence identity with 67-kDa calelectrin (p68), a protein with an 8-fold internal repeat. Comparing the amino-terminal domains of these four proteins of known sequence revealed that, in general, only endonexin II and protein II had significant sequence identity (29%). Endonexin II was not phosphorylated by Ca2+/phospholipid-dependent enzyme (protein kinase C) even though it contained a threonine at a position analogous to the protein kinase C phosphorylation sites of lipocortin I, calpactin I, and protein II.
Kaplan
R
R
Division of Molecular Biology, Rorer Biotechhology Inc., Springfield, Virginia 22151.
Jaye
M
M
Burgess
W H
WH
Schlaepfer
D D
DD
Haigler
H T
HT
D000595:Amino Acid Sequence; D017314:Annexin A4; D017304:Annexin A5; D017302:Annexins; D001483:Base Sequence; D002135:Calcium-Binding Proteins / Q000235:genetics* / Q000494:pharmacology*; D003001:Cloning, Molecular; D004247:DNA / Q000032:analysis; D006023:Glycoproteins / Q000494:pharmacology; D006801:Humans; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000235:genetics
D017314:Annexin A4; D017304:Annexin A5; D017302:Annexins; D002135:Calcium-Binding Proteins; D006023:Glycoproteins; D011994:Recombinant Proteins; D004247:DNA