Association of transglutaminase with the reconstituted keratin filaments isolated from rat vaginal epithelial cells.
We report a novel association of the calcium dependent cross-linking enzyme, transglutaminase (TGase) with the urea soluble reconstituted keratin filaments (RKF) isolated from the rat vaginal epithelial cells (VEC). This was ascertained by measuring the activity using 14C-spermidine incorporation and also by an increase in keratin filament aggregation by the addition of only TGase cofactor-calcium. These events were specifically inhibited by the treatment of calcium chelator, EDTA at a concentration > 2 mM as well as by pretreating the RKF with histamine, a TGase substrate inhibitor. The association was also exemplified by immunoblotting analysis where a specific and preferential polypeptide of molecular weight 58 kDa cross-reacted with TGase antibody amongst the other keratins. This phenomenon was not seen in the keratins isolated from skin, a non-targeting tissue for estradiol action.
Vijayalakshmi
V
V
Centre for Cellular and Molecular Biology, Hyderabad, India.
Gupta
P D
PD
D008841:Actin Cytoskeleton / Q000201:enzymology* / Q000648:ultrastructure; D000818:Animals; D004591:Electrophoresis, Polyacrylamide Gel; D004847:Epithelial Cells; D004848:Epithelium / Q000201:enzymology / Q000648:ultrastructure; D005260:Female; D015151:Immunoblotting; D007633:Keratins / Q000302:isolation & purification / Q000378:metabolism*; D007700:Kinetics; D009391:Nephelometry and Turbidimetry; D051381:Rats; D017208:Rats, Wistar; D011503:Transglutaminases / Q000302:isolation & purification / Q000378:metabolism*; D014621:Vagina / Q000166:cytology / Q000201:enzymology*
D007633:Keratins; D011503:Transglutaminases