Immunological characterization and fine localization of a lizard beta-keratin.
Scales of lizards contain beta-keratin of poorly known composition. In the present study, a rat polyclonal serum against a lizard beta-keratin of 14-15 kDa has been produced and the relative protein has been immunolocalized in the epidermis. The observations for the first time show that the isolated protein band derives from the extraction of a protein component of the beta-keratin filaments of lizard epidermis. In immunoblots and immunocytochemistry, the antiserum recognizes most lizard beta-keratins, but produces a variable cross-reactivity with snake beta-keratins, and weak or no reactivity with beta-keratins isolated from tuatara, turtles, alligator and birds. In bidimensional immunoblots of lizard epidermis, three main spots at 15-16 kDa with isoelectric point at 7.0, 7.6 and 8.0, and an unresolved large spot at 29-30 kDa and with pI at 7.5-8.0, are obtained, may be derived from the aggregation of smaller beta-keratin proteins. The ultrastructural immunolocalization with the antibody against lizard beta-keratin shows that only small and large beta-keratin filaments of beta-cells of lizard epidermis are labeled. Keratin bundles in oberhautchen cells are less immunolabeled. Beta-keratin is rapidly polymerized into beta-packets that merge into larger beta-keratin filaments. No labeling is present over other cell organelles or cell layers of lizard epidermis, and is absent in non-epidermal cells. The antiserum recognizes epitope(s) characteristics for lizard beta-keratins, partially recognized in snakes and absent in non-lepidosaurian species. This result indicates that beta-keratins among different reptilian groups posses different immunoreactive regions.
Alibardi
Lorenzo
L
Dipartimento di Biologia Evoluzionistica Sperimentale, University of Bologna, Bologna, Italy. alibardi@biblio.cib.unibo.it
Toni
Mattia
M
D000818:Animals; D004817:Epidermis / Q000737:chemistry*; D007106:Immune Sera / Q000276:immunology; D015151:Immunoblotting; D007150:Immunohistochemistry; D008116:Lizards / Q000276:immunology*; D051381:Rats; D013045:Species Specificity; D053800:beta-Keratins / Q000302:isolation & purification* / Q000648:ultrastructure
D007106:Immune Sera; D053800:beta-Keratins