Expression of transglutaminase 1 in human epidermis.
To explore the expression and function of the membrane-associated or type I transglutaminase (TGase1) in human epidermis, we have made a new antihuman TGase1 antibody in goats elicited against a purified active recombinant protein expressed in bacteria. By use of Western blotting and immunoprecipitation methods, the antibody reacted with high specificity with only the TGase1 activity of the epidermis and in cultured keratinocytes. By indirect immunofluorescence, the antibody decorated the entire epidermis, including the basal layer, with some potentiation of the granular layer. However, these staining properties are quite different from those of a widely used, commercially available TGase1 monoclonal antibody (termed B.C1), which decorates the granular layers of the epidermis. By Western blotting, it identifies the TGase1 protein band only weakly, but recognizes strongly a group of bands of 15-20 kDa, two of which by amino acid analysis and amino acid sequencing are the small proline-rich (SPR) 1 and SPR2 proteins, also expressed in epidermal and epithelial tissues. Together with a series of blocking experiments with TGase1 proteins and synthetic peptides, these data reveal that the major epitope of the B.C1 antibody most likely resides on the amino-terminus of these two SPR proteins rather than on TGase1. Further studies will now be necessary to determine the role(s) of TGase1 during the different stages of development and differentiation in the epidermis.
Kim
S Y
SY
Skin Biology Branch, National Institute of Arthritis and Musculoskeletal and Skin Diseases, National Institutes of Health, Bethesda, Maryland 20892.
Chung
S I
SI
Yoneda
K
K
Steinert
P M
PM
D000328:Adult; D000595:Amino Acid Sequence; D000906:Antibodies / Q000032:analysis / Q000276:immunology; D000911:Antibodies, Monoclonal; D000918:Antibody Specificity; D001667:Binding, Competitive / Q000187:drug effects; D055233:Cornified Envelope Proline-Rich Proteins; D000939:Epitopes / Q000187:drug effects; D005455:Fluorescent Antibody Technique; D006801:Humans; D008565:Membrane Proteins; D008969:Molecular Sequence Data; D010455:Peptides / Q000494:pharmacology; D011506:Proteins; D012333:RNA, Messenger / Q000032:analysis; D012867:Skin / Q000201:enzymology*; D011503:Transglutaminases / Q000235:genetics / Q000276:immunology* / Q000502:physiology*
D000906:Antibodies; D000911:Antibodies, Monoclonal; D055233:Cornified Envelope Proline-Rich Proteins; D000939:Epitopes; D008565:Membrane Proteins; D010455:Peptides; D011506:Proteins; D012333:RNA, Messenger; D011503:Transglutaminases