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Mass spectrometrical analysis of bilin‐binding protein from the wing of Hebomoia glaucippe (Linnaeus, 1758) (Lepidoptera: Pieridae)
Author(s) -
Bae Narkhyun,
Lödl Martin,
Pollak Arnold,
Lubec Gert
Publication year - 2012
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.201100569
Subject(s) - biochemistry , biology , chemistry , microbiology and biotechnology
Bilin‐binding protein (BBP) is a member of the lipocalin superfamily and a pigment protein in Lepidoptera. It is binding to a series of lipidic compounds but its functions remain to be elucidated. Working on wing proteins in H ebomoia glaucippe , we observed this protein on gels and decided to characterize BBP . A gel‐based mass spectrometrical method using two‐dimensional gel electrophoresis followed by in‐gel digestion of protein spots followed by nano‐ LC ‐ ESI ‐ MS / MS (ion trap, HCT ) identification and characterization of proteins was applied. An antibody was generated against the protein and immunoblotting in the butterfly and mouse brain was carried out. Two spots were identified from the butterfly wing as BBP ( P 09464) with high sequence coverage. Nitrotyrosination ( Y 163; as aminotyrosine) was observed and nitration was verified using immunoblotting. Additional posttranslational modifications (PTMs) as hypusine, carboxylation, kynurenine, aminoadipic acid, were proposed. The presence of BBP ‐immunoreactive protein was also observed in mouse brain. The characterization of BBP showed high sequence similarity with mouse apolipoprotein D and the findings suggest a tentative function of BBP comparable to apolipoproteins. The role of the PTMs remains elusive but nitration, in analogy to nitration effects reported in literature, proposes a role for mechanoelastic proteins and protein–protein interactions.