A major allergen in rainbow trout (Oncorhynchus mykiss): complete sequences of parvalbumin by MALDI tandem mass spectrometry.

نویسندگان

  • Donatella Aiello
  • Stefano Materazzi
  • Roberta Risoluti
  • Hariprasad Thangavel
  • Leonardo Di Donna
  • Fabio Mazzotti
  • Francesca Casadonte
  • Carlo Siciliano
  • Giovanni Sindona
  • Anna Napoli
چکیده

Fish parvalbumin (PRVB) is an abundant and stable protein in fish meat. The variation in cross-reactivity among individuals is well known and explained by a broad repertoire of molecular forms and differences between IgE-binding epitopes in fish species. PVRB has "sequential" epitopes, which retain their IgE-binding capacity and allergenicity also after heating and digestion using proteolytic enzymes. From the allergonomics perspective, PRVB is still a challenging target due to its multiple isoforms present at different degrees of distribution. Little information is available in the databases about PVRBs from Oncorhynchus mykiss. At present, only two validated, incomplete isoforms of this species are included in the protein databases: parvalbumin beta 1 (P86431) and parvalbumin beta 2 (P86432). A simple and rapid protocol has been developed for selective solubilization of PRVB from the muscle of farmed rainbow trout (Oncorhynchus mykiss), followed by calcium depletion, proteolytic digestion, MALDI MS, and MS/MS analysis. With this strategy thermal allergen release was assessed and PRVB1 (P86431), PRVB1.1, PRVB2 (P86432) and PRVB2.1 variants from the rainbow trout were sequenced. The correct ordering of peptide sequences was aided by mapping the overlapping enzymatic digests. The deduced peptide sequences were arranged and the theoretical molecular masses (Mr) of the resulting sequences were calculated. Experimental masses (Mr) of each PRVB variant were measured by linear MALDI-TOF.

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

منابع مشابه

A putative serine protease, SpSsp1, from Saprolegnia parasitica is recognised by sera of rainbow trout, Oncorhynchus mykiss

Saprolegniosis, the disease caused by Saprolegnia sp., results in considerable economic losses in aquaculture. Current control methods are inadequate, as they are either largely ineffective or present environmental and fish health concerns. Vaccination of fish presents an attractive alternative to these control methods. Therefore we set out to identify suitable antigens that could help generate...

متن کامل

Parvalbumin expression in trout swimming muscle correlates with relaxation rate.

Rainbow trout (Oncorhynchus mykiss) display longitudinal and developmental shifts in muscle relaxation rate. This study aimed to determine the role of variations in parvalbumin content in modulating muscle relaxation. Parvalbumin is a low molecular weight protein that buffers myoplasmic Ca2+ and enhances muscle relaxation. In some fish, longitudinal variations in muscle relaxation have been lin...

متن کامل

The influence of Wisconsinan glaciation and contemporary stream hydrology on microsatellite DNA variation in rainbow trout (Oncorhynchus mykiss)

Microsatellite DNA variation was examined in rainbow trout (Oncorhynchus mykiss) populations from throughout British Columbia, Canada, to address the roles of historical isolation, postglacial dispersal, and contemporary geomorphology in structuring genetic variation and differentiation. We detected signatures of historical isolation and postglacial recolonization in the form of ‘‘interior’’ an...

متن کامل

Cloning and sequencing of rainbow trout (Oncorhynchus mykiss) interferon regulatory factor 7

  Interferon regulatory factor 7 (IRF7) gene was cloned from a subtractive cDNA library constructed with mRNAs obtained from rainbow trout (Oncorhynchus mykiss) macrophage cell line (RTS-11). Using expressed sequence tag clones of submitted IRF7 amino acid sequences, specific primers were designed. Results showed that IRF7 cDNA contains an ORF of 1251 nucleotides that translates into a 416 resi...

متن کامل

ذخیره در منابع من


  با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

عنوان ژورنال:
  • Molecular bioSystems

دوره 11 8  شماره 

صفحات  -

تاریخ انتشار 2015