Molecular cloning and characterization of annexin V-binding proteins with highly hydrophilic peptide structure.
We previously reported that annexin V promoted the survival of cultured rat neocortical neurons. In an effort to elucidate the mechanism underlying this neurotrophic activity of annexin V, we have attempted to identify the target or binding proteins of annexin V in neuronal cells. Herein, we screened an embryonic day 17 rat brain cDNA library by western blot using glutathione S-transferase-annexin V fusion protein as a probe and then isolated four clones showing binding to annexin V in a Ca2+ - and phospholipid-dependent manner. Although these cDNAs encoded different polypeptides, all four polypeptides shared the unique feature of containing highly hydrophilic stretches with high Lys, Glu, and Ser contents. Deduced amino acid sequences of two clones showed high homology with human X-linked Helicase2 (XH2) and DNA (cytosine-5) methyltransferase (DMTase) sequences, whereas the other two were not related to any known peptide sequence. These results suggest that XH2 and DMTase are candidates for annexin V-binding proteins and thus may mediate the biological activity of annexin V.
Ohsawa
K
K
Department of Neurochemistry, National Institute of Neuroscience, Tokyo, Japan.
Imai
Y
Y
Ito
D
D
Kohsaka
S
S
D000818:Animals; D017304:Annexin A5 / Q000235:genetics / Q000378:metabolism*; D015153:Blotting, Western; D001921:Brain / Q000166:cytology / Q000502:physiology; D002352:Carrier Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D002467:Cell Nucleus / Q000737:chemistry; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D005333:Fetus / Q000502:physiology; D015723:Gene Library; D006801:Humans; D008969:Molecular Sequence Data; D009474:Neurons / Q000737:chemistry / Q000502:physiology; D010455:Peptides / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D051381:Rats; D017421:Sequence Analysis; D017386:Sequence Homology, Amino Acid
D017304:Annexin A5; D002352:Carrier Proteins; D018076:DNA, Complementary; D010455:Peptides