J. Biol. Chem., Vol. 259, Issue 13, 8037-8040, Jul, 1984

Synthetic peptides corresponding to keratin subunits elicit highly specific antibodies

DR Roop, CK Cheng, L Titterington, CA Meyers, JR Stanley, PM Steinert and SH Yuspa

The presence of common antigenic determinants within all keratin proteins has made difficult the production of antisera which are monospecific for individual keratin subunits. Synthetic peptides corresponding to the carboxyl-terminal amino acid sequences of the mouse 59- and 67-kilodalton keratins were used to produce antibodies which were highly specific for these keratin subunits. This method of antibody production was chosen after examination of amino acid sequence data (which were deduced from the nucleotide sequence of cDNA clones for these and other mouse keratins) revealed that the carboxyl-terminal amino acid sequences of various keratins were unique. Indirect immunofluorescence staining of newborn-mouse skin with these antisera demonstrated that the 59- and 67-kilodalton keratins were only present within the differentiated cells of the epidermis (the suprabasal layers) and not in the undifferentiated cells (the basal layer). These results are consistent with our previous work concerning the expression of these keratin genes at the messenger RNA level (Roop, D. R., Hawley- Nelson, P., Cheng, C. K., and Yuspa, S. H. (1983) Proc. Natl. Acad. Sci. U.S.A. 80, 716-720). Data obtained with the antisera directed against the 67-kilodalton keratin also indicated that the carboxyl- terminal sequences of this subunit were not present in the nonliving layer (the stratum corneum) of the epidermis. This approach should be useful for the production of antisera specific for other keratin subunits as additional sequence information becomes available.
CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				L. M. Sevilla, R. Nachat, K. R. Groot, J. F. Klement, J. Uitto, P. Djian, A. Maatta, and F. M. Watt Mice deficient in involucrin, envoplakin, and periplakin have a defective epidermal barrier J. Cell Biol., December 31, 2007; 179(7): 1599 - 1612. [Abstract] [Full Text] [PDF]

				J. Chen, X. Cheng, M. Merched-Sauvage, C. Caulin, D. R. Roop, and P. J. Koch An unexpected role for keratin 10 end domains in susceptibility to skin cancer J. Cell Sci., December 15, 2006; 119(24): 5067 - 5076. [Abstract] [Full Text] [PDF]

				Z. Du, K. Podsypanina, S. Huang, A. McGrath, M. J. Toneff, E. Bogoslovskaia, X. Zhang, R. C. Moraes, M. Fluck, D. C. Allred, et al. Introduction of oncogenes into mammary glands in vivo with an avian retroviral vector initiates and promotes carcinogenesis in mouse models PNAS, November 14, 2006; 103(46): 17396 - 17401. [Abstract] [Full Text] [PDF]

				C. Caulin, T. Nguyen, M. A. Longley, Z. Zhou, X.-J. Wang, and D. R. Roop Inducible Activation of Oncogenic K-ras Results in Tumor Formation in the Oral Cavity Cancer Res., August 1, 2004; 64(15): 5054 - 5058. [Abstract] [Full Text] [PDF]

				F. Benavides, M. F. Starost, M. Flores, I. B. Gimenez-Conti, J.-L. Guenet, and C. J. Conti Impaired Hair Follicle Morphogenesis and Cycling with Abnormal Epidermal Differentiation in nackt Mice, a Cathepsin L-Deficient Mutation Am. J. Pathol., August 1, 2002; 161(2): 693 - 703. [Abstract] [Full Text] [PDF]

				A. Hatoum, M. E. El-Sabban, J. Khoury, S. H. Yuspa, and N. Darwiche Overexpression of retinoic acid receptors alpha and gamma into neoplastic epidermal cells causes retinoic acid-induced growth arrest and apoptosis Carcinogenesis, December 1, 2001; 22(12): 1955 - 1963. [Abstract] [Full Text] [PDF]

				S. M. Wojcik, M. A. Longley, and D. R. Roop Discovery of a novel murine keratin 6 (K6) isoform explains the absence of hair and nail defects in mice deficient for K6a and K6b J. Cell Biol., August 6, 2001; 154(3): 619 - 630. [Abstract] [Full Text] [PDF]

				M. P. Santini, C. Talora, T. Seki, L. Bolgan, and G. P. Dotto Cross talk among calcineurin, Sp1/Sp3, and NFAT in control of p21WAF1/CIP1 expression in keratinocyte differentiation PNAS, August 1, 2001; (2001) 161299698. [Abstract] [Full Text] [PDF]

				M. J. Arin, M. A. Longley, X.-J. Wang, and D. R. Roop Focal Activation of a Mutant Allele Defines the Role of Stem Cells in Mosaic Skin Disorders J. Cell Biol., February 5, 2001; 152(3): 645 - 650. [Abstract] [Full Text] [PDF]

				P. J. Koch, P. A. de Viragh, E. Scharer, D. Bundman, M. A. Longley, J. Bickenbach, Y. Kawachi, Y. Suga, Z. Zhou, M. Huber, et al. Lessons from Loricrin-deficient Mice: Compensatory Mechanisms Maintaining Skin Barrier Function in the Absence of a Major Cornified Envelope Protein J. Cell Biol., October 18, 2000; 151(2): 389 - 400. [Abstract] [Full Text] [PDF]

				Y. Suga, M. Jarnik, P. S. Attar, M. A. Longley, D. Bundman, A. C. Steven, P. J. Koch, and D. R. Roop Transgenic Mice Expressing a Mutant Form of Loricrin Reveal the Molecular Basis of the Skin Diseases, Vohwinkel Syndrome and Progressive Symmetric Erythrokeratoderma J. Cell Biol., October 18, 2000; 151(2): 401 - 412. [Abstract] [Full Text] [PDF]

				K. M. Liefer, M. I. Koster, X.-J. Wang, A. Yang, F. McKeon, and D. R. Roop Down-Regulation of p63 Is Required for Epidermal UV-B-induced Apoptosis Cancer Res., August 1, 2000; 60(15): 4016 - 4020. [Abstract] [Full Text]

				S. M. Wojcik, D. S. Bundman, and D. R. Roop Delayed Wound Healing in Keratin 6a Knockout Mice Mol. Cell. Biol., July 15, 2000; 20(14): 5248 - 5255. [Abstract] [Full Text]

				D. B. Klug, E. Crouch, C. Carter, L. Coghlan, C. J. Conti, and E. R. Richie Transgenic Expression of Cyclin D1 in Thymic Epithelial Precursors Promotes Epithelial and T Cell Development J. Immunol., February 15, 2000; 164(4): 1881 - 1888. [Abstract] [Full Text] [PDF]

				P. M. Steinert and L. N. Marekov Initiation of Assembly of the Cell Envelope Barrier Structure of Stratified Squamous Epithelia Mol. Biol. Cell, December 1, 1999; 10(12): 4247 - 4261. [Abstract] [Full Text]

				D Danielpour Transdifferentiation of NRP-152 rat prostatic basal epithelial cells toward a luminal phenotype: regulation by glucocorticoid, insulin-like growth factor-I and transforming growth factor-beta J. Cell Sci., January 1, 1999; 112(2): 169 - 179. [Abstract] [PDF]

				T. N. Seagroves, S. Krnacik, B. Raught, J. Gay, B. Burgess-Beusse, G. J. Darlington, and J. M. Rosen C/EBPbeta , but not C/EBPalpha , is essential for ductal morphogenesis, lobuloalveolar proliferation, and functional differentiation in the mouse mammary gland Genes & Dev., June 15, 1998; 12(12): 1917 - 1928. [Abstract] [Full Text]

				B Andersen, W C Weinberg, O Rennekampff, R J McEvilly, J R Bermingham, F Hooshmand, V Vasilyev, J F Hansbrough, M R Pittelkow, S H Yuspa, et al. Functions of the POU domain genes Skn-1a/i and Tst-1/Oct-6/SCIP in epidermal differentiation. Genes & Dev., July 15, 1997; 11(14): 1873 - 1884. [Abstract] [PDF]

				K. Takahashi and P. A. Coulombe Defining a Region of the Human Keratin 6a Gene That Confers Inducible Expression in Stratified Epithelia of Transgenic Mice J. Biol. Chem., May 2, 1997; 272(18): 11979 - 11985. [Abstract] [Full Text] [PDF]

				P. M. Steinert and L. N. Marekov Direct Evidence That Involucrin Is a Major Early Isopeptide Cross-linked Component of the Keratinocyte Cornified Cell Envelope J. Biol. Chem., January 17, 1997; 272(3): 2021 - 2030. [Abstract] [Full Text] [PDF]

				J. Paramio, M. Casanova, A Alonso, and J. Jorcano Keratin intermediate filament dynamics in cell heterokaryons reveals diverse behaviour of different keratins J. Cell Sci., January 5, 1997; 110(9): 1099 - 1111. [Abstract] [PDF]

				K. Takahashi and P. A. Coulombe A transgenic mouse model with an inducible skin blistering disease phenotype PNAS, December 10, 1996; 93(25): 14776 - 14781. [Abstract] [Full Text] [PDF]

				M. P. Santini, C. Talora, T. Seki, L. Bolgan, and G. P. Dotto Cross talk among calcineurin, Sp1/Sp3, and NFAT in control of p21WAF1/CIP1 expression in keratinocyte differentiation PNAS, August 14, 2001; 98(17): 9575 - 9580. [Abstract] [Full Text] [PDF]