Identification of a preferred substrate peptide for transglutaminase 3 and detection of in situ activity in skin and hair follicles.
Transglutaminases (TGases) are a family of enzymes that catalyze cross-linking reactions between proteins. During epidermal differentiation, these enzymatic reactions are essential for formation of the cornified envelope, which consists of cross-linked structural proteins. Two main transglutaminases isoforms, epidermal-type (TGase 3) and keratinocyte-type (TGase 1), are cooperatively involved in this process of differentiating keratinocytes. Information regarding their substrate preference is of great importance to determine the functional role of these isozymes and clarify their possible co-operative action. Thus far, we have identified highly reactive peptide sequences specifically recognized by TGases isozymes such as TGase 1, TGase 2 (tissue-type isozyme) and the blood coagulation isozyme, Factor XIII. In this study, several substrate peptide sequences for human TGase 3 were screened from a phage-displayed peptide library. The preferred substrate sequences for TGase 3 were selected and evaluated as fusion proteins with mutated glutathione S-transferase. From these studies, a highly reactive and isozyme-specific sequence (E51) was identified. Furthermore, this sequence was found to be a prominent substrate in the peptide form and was suitable for detection of in situ TGase 3 activity in the mouse epidermis. TGase 3 enzymatic activity was detected in the layers of differentiating keratinocytes and hair follicles with patterns distinct from those of TGase 1. Our findings provide new information on the specific distribution of TGase 3 and constitute a useful tool to clarify its functional role in the epidermis.
Yamane
Asaka
A
Department of Applied Molecular Biosciences, Graduate School of Bioagricultural Sciences, Nagoya University, Japan.
Fukui
Mina
M
Sugimura
Yoshiaki
Y
Itoh
Miho
M
Alea
Mileidys Perez
MP
Thomas
Vincent
V
El Alaoui
Said
S
Akiyama
Masashi
M
Hitomi
Kiyotaka
K
D000595:Amino Acid Sequence; D000818:Animals; D018859:Hair Follicle / Q000201:enzymology*; D006801:Humans; D007527:Isoenzymes / Q000378:metabolism; D051379:Mice; D008969:Molecular Sequence Data; D019151:Peptide Library; D010455:Peptides / Q000737:chemistry / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism; D012867:Skin / Q000201:enzymology*; D013379:Substrate Specificity; D011503:Transglutaminases / Q000378:metabolism*
D007527:Isoenzymes; D019151:Peptide Library; D010455:Peptides; D011994:Recombinant Proteins; C498713:Tgm3 protein, mouse; C498712:TGM3 protein, human; D011503:Transglutaminases