Isoform-specific intracellular vesicle formation by recombinant annexin XI-A in Sf9 cells.
Annexins are a group of structurally related proteins that bind phospholipids in a Ca2(+)-dependent manner and have the ability to self-aggregate and to promote vesicle aggregation and membrane fusion. Two isoforms of annexin XI, termed XI-A and XI-B, were previously identified by screening a bovine chondrocyte cDNA library. But little is known about differences in their biological function. In the present study, we therefore examined the results of expression of the two proteins in Spodoptera frugiperda (Sf9) insect cells, and in mammalian COS-7 cells. Annexin XI isoforms were expressed in Sf9 cells using a baculovirus expression system. Recombinant annexin XI-A but not XI-B caused formation of spherical "annexin XI-associated vesicles, " in the cytoplasm of Sf9 cells. Furthermore, indirect immnocytofluorescence studies showed similar phenomenon, that of local aggregation, with transfected annexin XI-A in COS-7 cells, whereas annexin XI-B remained diffusely distributed throughout the cytoplasm. Since annexin XI isoforms differ in amino acid sequence only in the alternative splicing region of the N-terminal domain, these findings suggest that this domain has distinct biological significance in terms of aggregation and vesicle formation.
Sudo
T
T
Department of Pharmacology, Nagoya University School of Medicine, Japan.
Mamiya
N
N
Goto
M
M
Watanabe
Y
Y
Hidaka
H
H
D000595:Amino Acid Sequence; D000818:Animals; D017302:Annexins / Q000096:biosynthesis* / Q000737:chemistry / Q000302:isolation & purification; D002356:Cartilage / Q000378:metabolism; D002417:Cattle; D002460:Cell Line; D002522:Chlorocebus aethiops; D018076:DNA, Complementary; D015723:Gene Library; D014644:Genetic Variation; D008969:Molecular Sequence Data; D015388:Organelles / Q000378:metabolism / Q000648:ultrastructure; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification; D018411:Spodoptera; D014162:Transfection
D017302:Annexins; D018076:DNA, Complementary; D011994:Recombinant Proteins