Identification of a lectin protein from black turtle bean (Phaseolus vulgaris) using LC-MS/MS and PCR method.
He, S.-D., Shi, J., Li, X., Ma, Y., and Xue, S.J. (2015). "Identification of a lectin protein from black turtle bean (Phaseolus vulgaris) using LC-MS/MS and PCR method.", Food Science and Technology - LWT, 60(2, Part 2), pp. 1074-1079. doi : 10.1016/j.lwt.2014.10.015 Access to full text
A 31 KDa lectin isolated from the black turtle bean (Phaseolus vulgaris) was identified by mass spectrometry, and the DNA fragment coding for the lectin was elucidated by the cloning and sequence analysis. The protein faction was separated by SDS-PAGE and was digested with trypsin. Then the peptide fragments were analyzed by LC-MS/MS. De novo sequencing software was employed to identify homologous peptides against the NCBI database using the MS-BLAST searching method. Based on the MS results, the coding sequence of the lectin was obtained by PCR method. The sequence analysis was done using Clustal W2 and MEGA 5. MS analysis of the protein was 52% sequenced and the protein was identified as a lectin with a strong homology of phytohemagglutinin (PHA) genes. The full-length cDNA of the lectin (KF601826) with 934 bp was amplified, containing one main highly conserved domain of lectin_legume_LecRK_Arcelin_ConA region. Multiple alignment and phylogenetic analysis clearly indicated that the lectin belonged to the PHA family, and the lectin might evolve from the Arc7 variant. Results obtained can serve as a base for future studies on food safety control and biological properties of the lectin.
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