RRC ID 64809
著者 Jang YR, Cho K, Kim SW, Altenbach SB, Lim SH, Sim JR, Lee JY.
タイトル Development of an Optimized MALDI-TOF-MS Method for High-Throughput Identification of High-Molecular-Weight Glutenin Subunits in Wheat.
ジャーナル Molecules
Abstract Because high-molecular-weight glutenin subunits (HMW-GS) are important contributors to wheat end-use quality, there is a need for high-throughput identification of HMW-GS in wheat genetic resources and breeding lines. We developed an optimized method using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) to distinguish individual HMW-GS by considering the effects of the alkylating reagent in protein extraction, solvent components, dissolving volume, and matrix II components. Using the optimized method, 18 of 22 HMW-GS were successfully identified in standard wheat cultivars by differences in molecular weights or by their associations with other tightly linked subunits. Interestingly, 1Bx7 subunits were divided into 1Bx7 group 1 and 1Bx7 group 2 proteins with molecular weights of about 82,400 and 83,000 Da, respectively. Cultivars containing the 1Bx7 group 2 proteins were distinguished from those containing 1Bx7OE using well-known DNA markers. HMW-GS 1Ax2* and 1Bx6 and 1By8 and 1By8*, which are difficult to distinguish due to very similar molecular weights, were easily identified using RP-HPLC. To validate the method, HMW-GS from 38 Korean wheat varieties previously evaluated by SDS-PAGE combined with RP-HPLC were analyzed by MALDI-TOF-MS. The optimized MALDI-TOF-MS method will be a rapid, high-throughput tool for selecting lines containing desirable HMW-GS for breeding efforts.
巻・号 25(18)
公開日 2020-9-22
DOI 10.3390/molecules25184347
PII molecules25184347
PMID 32971947
PMC PMC7571055
MeSH Glutens / analysis* Glutens / chemistry* Molecular Weight Protein Subunits / analysis* Protein Subunits / chemistry* Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / methods* Triticum / chemistry*
IF 3.267
リソース情報
コムギ Chinese Spring