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Development of an Optimized MALDI-TOF-MS Method for High-Throughput Identification of High-Molecular-Weight Glutenin Subunits in Wheat

1
National Institute of Agricultural Science, RDA, Jeonju 54874, Korea
2
Department of Biotechnology, College of Agriculture and Life Sciences, Chonnam National University, Gwangju 500-757, Korea
3
USDA-ARS, Western Regional Research Center, 800 Buchanan Street, Albany, CA 94710, USA
4
Division of Horticultural Biotechnology, Hankyong National University, Anseong 17579, Korea
*
Author to whom correspondence should be addressed.
These authors contributed equally to this work.
Academic Editor: Daniel Cozzolino
Molecules 2020, 25(18), 4347; https://doi.org/10.3390/molecules25184347
Received: 1 September 2020 / Revised: 17 September 2020 / Accepted: 17 September 2020 / Published: 22 September 2020
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. View Full-Text
Keywords: high-molecular-weight glutenin subunits; MALDI-TOF-MS; wheat flour quality high-molecular-weight glutenin subunits; MALDI-TOF-MS; wheat flour quality
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MDPI and ACS Style

Jang, Y.-R.; Cho, K.; Kim, S.W.; Altenbach, S.B.; Lim, S.-H.; Sim, J.-R.; Lee, J.-Y. Development of an Optimized MALDI-TOF-MS Method for High-Throughput Identification of High-Molecular-Weight Glutenin Subunits in Wheat. Molecules 2020, 25, 4347. https://doi.org/10.3390/molecules25184347

AMA Style

Jang Y-R, Cho K, Kim SW, Altenbach SB, Lim S-H, Sim J-R, Lee J-Y. Development of an Optimized MALDI-TOF-MS Method for High-Throughput Identification of High-Molecular-Weight Glutenin Subunits in Wheat. Molecules. 2020; 25(18):4347. https://doi.org/10.3390/molecules25184347

Chicago/Turabian Style

Jang, You-Ran, Kyoungwon Cho, Se W. Kim, Susan B. Altenbach, Sun-Hyung Lim, Jae-Ryeong Sim, and Jong-Yeol Lee. 2020. "Development of an Optimized MALDI-TOF-MS Method for High-Throughput Identification of High-Molecular-Weight Glutenin Subunits in Wheat" Molecules 25, no. 18: 4347. https://doi.org/10.3390/molecules25184347

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