title: Recombinant human tissue transglutaminase produced into tobacco suspension cell cultures is active and recognizes autoantibodies in the serum of coeliac patients.
abstract: Human tissue transglutaminase (htTG) is one of the most important member within the transglutaminase family, enzymes that for their capacity of catalyzing post-translational modifications of proteins and peptides, rise an high interest for industrial applications. More recently, for its implication as the major autoantigen in the coeliac disease, availability of human tissue transglutaminase as recombinant form is required for accurate diagnostic tests. The aim of this study was to find an alternative and inexpensive source to produce human tissue transglutaminase. To date, plant systems are proposed as heterologous hosts to produce recombinant proteins for use in disease diagnosis and therapy. Here, we describe the stable expression of human tissue transglutaminase into Nicotiana tabacum cultured cells (cultivar Bright Yellow 2 (BY-2)). The recombinant enzyme was successfully expressed in different plant cell compartments and both apoplast (apo) and chloroplast (chl) purified proteins were shown to be catalytically active and able to bind GTP, a property possessed by the natural counterpart. Importantly, plant produced human tissue transglutaminase recognized autoantibodies in the serum of coeliac patients, suggesting possible applications in the diagnosis of coeliac disease.
authors.affiliation: Department of Food Science, University of Naples "Federico II", Parco Gussone, Portici 80055, Naples, Italy.
authors.forename: A
authors.initials: A
authors.lastname: Sorrentino
authors.forename: S
authors.initials: S
authors.lastname: Schillberg
authors.forename: R
authors.initials: R
authors.lastname: Fischer
authors.forename: R
authors.initials: R
authors.lastname: Rao
authors.forename: R
authors.initials: R
authors.lastname: Porta
authors.forename: L
authors.initials: L
authors.lastname: Mariniello
mesh_terms: D001323:Autoantibodies / Q000097:blood / Q000276:immunology*; D001483:Base Sequence; D002446:Celiac Disease / Q000097:blood*; D018929:Cell Culture Techniques; D002846:Chromatography, Affinity; D017931:DNA Primers; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D006160:Guanosine Triphosphate / Q000378:metabolism; D006801:Humans; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism; D014026:Nicotiana / Q000378:metabolism*; D011503:Transglutaminases / Q000096:biosynthesis* / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism
chemical_list: D001323:Autoantibodies; D017931:DNA Primers; D011994:Recombinant Proteins; D006160:Guanosine Triphosphate; D011503:Transglutaminases