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Top-down HPLC-ESI-MS characterization of rat gliadoralin A, a new member of the family of rat submandibular gland glutamine-rich proteins and potential substrate of transglutaminase

Cabras, Tiziana ; Iavarone, Federica ; Pirolli, Davide ; Cristina De Rosa, Maria ; Vitali, Alberto ; Faa, Gavino ; Cordaro, Massimo ; Messana, Irene ; Ekström, Jörgen ; Castagnola, Massimo

Journal of separation science, 2013-09, Vol.36 (17), p.2848-2861 [Periódico revisado por pares]

Weinheim: Blackwell Publishing Ltd

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Título:
Top-down HPLC-ESI-MS characterization of rat gliadoralin A, a new member of the family of rat submandibular gland glutamine-rich proteins and potential substrate of transglutaminase
Autor: Cabras, Tiziana ; Iavarone, Federica ; Pirolli, Davide ; Cristina De Rosa, Maria ; Vitali, Alberto ; Faa, Gavino ; Cordaro, Massimo ; Messana, Irene ; Ekström, Jörgen ; Castagnola, Massimo
Assuntos: 1987 ; 1990 ; Amino acids ; Analytical, structural and metabolic biochemistry ; Animals ; ARCHIVES OF ORAL BIOLOGY ; ATES OF AMERICA ; Biological and medical sciences ; Chemical Sciences ; Chromatography, High Pressure Liquid - methods ; CROSS-LINKING ; DIFFERENT PROTEOMIC PLATFORMS ; Enzymes ; EXPRESSION ; Fragmentation ; Fundamental and applied biological sciences. Psychology ; Gene sequencing ; General aspects, investigation methods ; Glutamine ; Glutamine - chemistry ; Glutamine-rich protein ; HUMAN ; JOURNAL OF BIOLOGICAL CHEMISTRY ; Kemi ; MASS-SPECTROMETRY ; P7289 ; P8472 ; PELLICLE ; PEPTIDES ; Proteins ; Rat ; Rats ; RELS L ; Residues ; Rodents ; Saliva ; Salivary Proteins and Peptides - chemistry ; Salivary Proteins and Peptides - metabolism ; SALIVARY PROTEOME ; SECONDARY STRUCTURE PREDICTION ; SERVER ; Spectrometry, Mass, Electrospray Ionization - methods ; Stimulation ; Submandibular ; Submandibular Gland - chemistry ; Substrate Specificity ; Substrates ; TISSUE ; Top-down ; TRANSGLUTAMINASE ; Transglutaminases - metabolism ; V262 ; V35 ; V87
É parte de: Journal of separation science, 2013-09, Vol.36 (17), p.2848-2861
Notas: Cagliari University
ArticleID:JSSC3308
istex:C1184B2A57B0379FEA3DEF48A07BB5F7F403E977
Regione Sardegna and Nando Peretti Foundation
ark:/67375/WNG-D3NGNR7T-9
Catholic University of Rome
Italian National Research Council (CNR)
MIUR
See the article online to view Fig. 7 in colour.
This article is dedicated to Frantisek Svec on the occasion of his 70th birthday.
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Descrição: During HPLC–ESI‐MS/MS analysis of rat submandibular saliva secreted under isoprenaline stimulation, a protein with an experimental [M+H]1+ = 10 544.24 m/z was detected (17.5 ± 0.7 min). The MS/MS fragmentation pattern, manually investigated, allowed establishing an internal sequence in agreement with a DNA‐derived sequence of an unknown rat protein coded D3Z9M3 (Swiss‐Prot). To match the experimental MS/MS fragmentation pattern and protein mass with theoretical data, the removal from the N terminus of the signal peptide and from the C terminus of three amino acid (a.a.) residues (Arg‐Ala‐Val) and the cyclization of the N‐terminal glutamine in pyroglutamic had to be supposed, resulting in a mature protein of 90 a.a. HPLC–ESI‐MS/MS of the trypsin digest ensured 100% sequence coverage. For the high glutamine content (34/90 = 37.8%) we propose to name this protein rat gliadoralin A 1–90. Low amounts of five different isoforms were sporadically detected, which did not significantly change their relative amounts after stimulation. Gliadoralin A is substrate for transglutaminase‐2, having Lys 60 and different Gln residues as major determinants for enzyme recognition. In silico investigation of superior structures evidenced that a small part of the protein adopts an α‐helical fold, whereas large segments are unfolded, suggesting an unordered conformation.
Editor: Weinheim: Blackwell Publishing Ltd
Idioma: Inglês

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Top-down HPLC-ESI-MS characterization of rat gliadoralin A, a new member of the family of rat submandibular gland glutamine-rich proteins and potential substrate of transglutaminase

Cabras, Tiziana ; Iavarone, Federica ; Pirolli, Davide ; Cristina De Rosa, Maria ; Vitali, Alberto ; Faa, Gavino ; Cordaro, Massimo ; Messana, Irene ; Ekström, Jörgen ; Castagnola, Massimo

Weinheim: Blackwell Publishing Ltd

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