title: Glutenase and collagenase activities of wheat cysteine protease Triticain-α: feasibility for enzymatic therapy assays.
abstract: Insufficient and/or improper protein degradation is associated with the development of various human pathologies. Enzymatic therapy with proteolytic enzymes aimed to improve insufficient proteolytic activity was suggested as a treatment of protease deficiency-induced disorders. Since in many cases human degradome is incapable of degrading the entire target protein(s), other organisms can be used as a source of proteases exhibiting activities distinct from human enzymes, and plants are perspective candidates for this source. In this study recombinant wheat cysteine protease Triticain-α was shown to refold in vitro into an autocatalytically activated proteolytic enzyme possessing glutenase and collagenase activities at acidic (or close to neutral) pH levels at the temperature of human body. Mass-spectrometry analysis of the products of Triticain-α-catalyzed gluten hydrolysis revealed multiple cleavage sites within the sequences of gliadin toxic peptides, in particular, in the major toxic 33-mer α-gliadin-derived peptide initiating inflammatory responses to gluten in celiac disease (CD) patients. Triticain-α was found to be relatively stable in the conditions simulating stomach environment. We conclude that Triticain-α can be exploited as a basic compound for development of (i) pharmaceuticals for oral administration aimed at release of the active enzyme into the gastric lumen for CD treatment, and (ii) topically active pharmaceuticals for wound debridement applications.
authors.affiliation: Sechenov First Moscow State Medical University, Institute of Molecular Medicine, 119991, Moscow, Russia.
authors.forename: Lyudmila V
authors.initials: LV
authors.lastname: Savvateeva
authors.affiliation: Sechenov First Moscow State Medical University, Institute of Molecular Medicine, 119991, Moscow, Russia.
authors.forename: Neonila V
authors.initials: NV
authors.lastname: Gorokhovets
authors.affiliation: Sechenov First Moscow State Medical University, Institute of Molecular Medicine, 119991, Moscow, Russia.
authors.forename: Vladimir A
authors.initials: VA
authors.lastname: Makarov
authors.affiliation: Lomonosov Moscow State University, Belozersky Institute of Physico-Chemical Biology, 119991, Moscow, Russia.
authors.forename: Marina V
authors.initials: MV
authors.lastname: Serebryakova
authors.affiliation: Lomonosov Moscow State University, Belozersky Institute of Physico-Chemical Biology, 119991, Moscow, Russia.
authors.forename: Andrey G
authors.initials: AG
authors.lastname: Solovyev
authors.affiliation: Lomonosov Moscow State University, Belozersky Institute of Physico-Chemical Biology, 119991, Moscow, Russia.
authors.forename: Sergey Yu
authors.initials: SY
authors.lastname: Morozov
authors.affiliation: Missouri University of Science and Technology, Rolla, MO, 65409, USA.
authors.forename: V Prakash
authors.initials: VP
authors.lastname: Reddy
authors.affiliation: Lomonosov Moscow State University, Belozersky Institute of Physico-Chemical Biology, 119991, Moscow, Russia.
authors.forename: Evgeni Yu
authors.initials: EY
authors.lastname: Zernii
authors.affiliation: Sechenov First Moscow State Medical University, Institute of Molecular Medicine, 119991, Moscow, Russia; Lomonosov Moscow State University, Belozersky Institute of Physico-Chemical Biology, 119991, Moscow, Russia. Electronic address: zamyat@belozersky.msu.ru.
authors.forename: Andrey A
authors.initials: AA
authors.lastname: Zamyatnin
authors.affiliation: "GALLY" International Biomedical Research Consulting LLC, San Antonio, TX, 78229, USA; School of Health Science and Healthcare Administration, The University of Atlanta, Johns Creek, GA, 30097, USA. Electronic address: GAliev@uofa.edu.
authors.forename: Gjumrakch
authors.initials: G
authors.lastname: Aliev
mesh_terms: D000595:Amino Acid Sequence; D002446:Celiac Disease / Q000188:drug therapy; D017364:Collagenases / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism* / Q000627:therapeutic use; D057056:Cysteine Proteases / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism* / Q000627:therapeutic use; D003646:Debridement / Q000379:methods; D056947:Enzyme Replacement Therapy; D005240:Feasibility Studies; D005983:Glutens / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism* / Q000627:therapeutic use; D006801:Humans; D008969:Molecular Sequence Data; D010940:Plant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism* / Q000627:therapeutic use; D059748:Proteolysis; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism / Q000627:therapeutic use; D014908:Triticum / Q000201:enzymology* / Q000235:genetics
chemical_list: D010940:Plant Proteins; D011994:Recombinant Proteins; D005983:Glutens; D057056:Cysteine Proteases; C000606744:triticain-alpha protein, Triticum aestivum; D017364:Collagenases
keywords: Collagen; Gliadin; Glutenin; Protease; Proteolytic cleavage