Retinol-induced morphological changes of cultured bovine endothelial cells are accompanied by a marked increase in transglutaminase.
Retinol, a morphogen, has been shown to induce morphological changes in vascular endothelial cells, accompanied by an acute and specific accumulation of an 80-kDa protein; purification and characterization of this retinol-induced protein (RIP) have revealed that it is a transglutaminase. Endothelial cells from bovine carotid artery were cultured, treated with retinol, and examined for changes in morphology and protein profiles. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of extracts prepared from retinol-treated cells which had undergone a remarkable change in shape (from a cobblestone-like to a spindle-like shape) indicated that the retinol-induced morphological change is accompanied by a marked increase of an 80-kDa protein. Similar changes were also induced by retinoic acid. The 80-kDa RIP was purified by anion exchange and hydroxyapatite column chromatography. Amino acid sequencing of tryptic fragments of the purified RIP revealed a high degree of homology between the sequence of bovine RIP and that of guinea pig liver transglutaminase, suggesting that RIP is a transglutaminase. This was confirmed by activity measurements; RIP exhibited transglutaminase activity, and an antiserum against RIP immunoprecipitated the activity. These results suggest that transglutaminase plays important roles in the maintenance of morphology and the control of endothelial cell functions.
Nara
K
K
Department of Biological Sciences, Tokyo Institute of Technology, Japan.
Nakanishi
K
K
Hagiwara
H
H
Wakita
K
K
Kojima
S
S
Hirose
S
S
D000595:Amino Acid Sequence; D000818:Animals; D002339:Carotid Arteries; D002417:Cattle; D002478:Cells, Cultured; D002851:Chromatography, High Pressure Liquid; D003600:Cytosol / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects / Q000201:enzymology*; D004790:Enzyme Induction; D015151:Immunoblotting; D007700:Kinetics; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010446:Peptide Fragments / Q000302:isolation & purification; D011503:Transglutaminases / Q000096:biosynthesis* / Q000302:isolation & purification; D014357:Trypsin; D014801:Vitamin A / Q000494:pharmacology*
D010446:Peptide Fragments; D014801:Vitamin A; D011503:Transglutaminases; D014357:Trypsin