A keratin antibody recognizing a heterotypic complex: epitope mapping to complementary locations on both components of the complex

Experimental Cell Research
A WaseemN H Waseem

Abstract

Keratin filaments in simple epithelial cells are heteropolymers of keratin 8 (K8) and keratin 18 (K18), which can be stained by the monoclonal antibody (MAb) LE61. This antibody has been widely used to study keratin expression in normal and neoplastic tissues. In this study we have found that MAb LE61 does not react with individual keratin polypeptides either derived from natural sources or expressed as recombinant proteins in Escherichia coli. However, when K8 or K18 bound to nitrocellulose were incubated with complementary keratin they became reactive with this antibody. A mixture of K8 and K18 in solution also reacted strongly with the MAb LE61 in ELISA. These observations suggest that the antibody recognizes a discontinuous epitope on the keratin complex. The antibody also reacted with complexes of K8 and K18 with other keratins. To locate the epitope of this antibody we have expressed K8 and K18 fragments, deleted from the amino- and carboxyl-termini, as fusion proteins with glutathione S-transferase. These fragments were able to form a heterotypic complex with the complementary keratin. Binding of the MAb LE61 to these complexes mapped the two halves of the epitope on K8, between residues 353 and 367, and on K18, between ...Continue Reading

Citations

Apr 2, 2002·Neoplasia : an International Journal for Oncology Research·Stephen MitchellEl-Nasir Lalani
Jan 22, 2008·Journal of Oral Pathology & Medicine : Official Publication of the International Association of Oral Pathologists and the American Academy of Oral Pathology·Anand LalliAhmad Waseem
Jun 20, 2007·Experimental Dermatology·Baki AkgülAlan Storey
Jun 15, 2004·Experimental Cell Research·Bert SchutteFrans C S Ramaekers
Mar 20, 1999·The Journal of Investigative Dermatology·A WaseemI M Leigh
Mar 30, 2002·The Journal of Biological Chemistry·Henrik J DitzelJens C Jensenius
Jul 1, 1997·The International Journal of Biochemistry & Cell Biology·A WaseemN H Waseem

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