Using mass spectrometry to detect buffalo salivary odorant-binding protein and its post-translational modifications.
A large number of mammalian odorant-binding proteins, which are lipocalins, have been studied. These proteins participate in peri-receptor events by selecting and carrying odorant molecules. The present study aimed at identifying the buffalo salivary odorant-binding protein (sOBP), and to determine its post-translational modification using mass spectrometry. The buffalo salivary 21  kDa protein was initially separated adopting sodium dodecyl sulfate-polyacrylamide gel electrophoresis and it was identified as sOBP with high statistical reliability using liquid chromatography/tandem mass spectrometry (LC/MS/MS) and SEQUEST, for the first time. Further, the post-translationally modified peptides were screened adopting MS/MS. A total of four post-translational modifications, namely glycation at lysine-(59), hydroxylation at lysine-(134), ubiquitination at lysine-(121), and dihydroxylation in lysine-(108), were recorded. Moreover, these modifications have not been identified in buffalo salivary odorant-binding protein.
Rajkumar
R
R
Center for Pheromone Technology, Department of Animal Science, Bharathidasan University, Trichirappalli 620 024, Tamilnadu, India.
Karthikeyan
K
K
Archunan
G
G
Huang
P H
PH
Chen
Y W
YW
Ng
W V
WV
Liao
C C
CC
D000595:Amino Acid Sequence; D000818:Animals; D002020:Buffaloes; D002853:Chromatography, Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D006023:Glycoproteins / Q000737:chemistry / Q000378:metabolism; D006031:Glycosylation; D006900:Hydroxylation; D008239:Lysine / Q000737:chemistry / Q000378:metabolism; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010503:Periodic Acid-Schiff Reaction; D011499:Protein Processing, Post-Translational; D017434:Protein Structure, Tertiary; D018035:Receptors, Odorant / Q000737:chemistry / Q000378:metabolism*; D012471:Salivary Proteins and Peptides / Q000737:chemistry / Q000378:metabolism*; D016415:Sequence Alignment; D053719:Tandem Mass Spectrometry / Q000379:methods*; D054875:Ubiquitination
D006023:Glycoproteins; D018035:Receptors, Odorant; D012471:Salivary Proteins and Peptides; D008239:Lysine