title: Transglutaminase1 preferred substrate peptide K5 is an efficient tool in diagnosis of lamellar ichthyosis.
abstract: Lamellar ichthyosis (LI) is a genetically heterogeneous, severe genodermatosis showing widespread hyperkeratosis of the skin. Transglutaminase 1 (TGase1) deficiency by TGase1 gene (TGM1) mutations is the most prevalent cause of LI. Screening of TGase1 deficiency in skin is essential to facilitate the molecular diagnosis of LI. However, cadaverine, the most widely used substrate for TGase activity assay, is not isozyme specific. Recently, a human TGase1-specific highly preferred substrate peptide K5 (pepK5) was generated. To evaluate its potential as a diagnostic tool for LI, we performed pepK5 labeling of TGase1 activity in normal human and LI skin. Ca(2+)-dependent labeling of FITC-pepK5 was clearly seen in the upper spinous and granular layers of normal human skin where it precisely overlapped with TGase1 immunostaining. Both specificity and sensitivity of FITC-pepK5 labeling for TGase1 activity were higher than those of FITC-cadaverine labeling. FITC-pepK5 labeling colocalized with involucrin and loricrin immunostaining at cornified cell envelope forming sites. FITC-pepK5 labeling was negative in LI patients carrying TGM1 truncation mutations and partially abolished in the other LI patients harboring missense mutations. The present results clearly indicate that pepK5 is a powerful tool for screening LI patient TGase1 deficiency when we make molecular diagnosis of LI.
authors.affiliation: Department of Dermatology, Hokkaido University Graduate School of Medicine, Sapporo 060-8638, Japan. akiyama@med.hokudai.ac.jp
authors.forename: Masashi
authors.initials: M
authors.lastname: Akiyama
authors.forename: Kaori
authors.initials: K
authors.lastname: Sakai
authors.forename: Teruki
authors.initials: T
authors.lastname: Yanagi
authors.forename: Satoshi
authors.initials: S
authors.lastname: Fukushima
authors.forename: Hironobu
authors.initials: H
authors.lastname: Ihn
authors.forename: Kiyotaka
authors.initials: K
authors.lastname: Hitomi
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Shimizu
mesh_terms: D001706:Biopsy; D002103:Cadaverine / Q000737:chemistry; D016022:Case-Control Studies; D004252:DNA Mutational Analysis; D006801:Humans; D017490:Ichthyosis, Lamellar / Q000175:diagnosis* / Q000378:metabolism*; D007527:Isoenzymes / Q000737:chemistry; D008856:Microscopy, Fluorescence / Q000379:methods; D009154:Mutation; D010455:Peptides / Q000737:chemistry*; D010641:Phenotype; D012867:Skin / Q000473:pathology; D013379:Substrate Specificity; D011503:Transglutaminases / Q000378:metabolism / Q000502:physiology*
chemical_list: D007527:Isoenzymes; C083605:K5 peptide; D010455:Peptides; D011503:Transglutaminases; C105804:transglutaminase 1; D002103:Cadaverine