Proteomics

Dataset Information

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Quantitation of ATIs in Wheat by LC-MS/MS – LFQ Orbitrap

ABSTRACT: A selection of ATIs was silenced in the common wheat cultivar Bobwhite and durum wheat cultivar Svevo by RNAi and gene editing, respectively, in order to reduce the amounts of ATIs. The controls and silenced lines were analyzed after digestion to peptides by LC-MS/MS with different approaches to evaluate changes in composition of ATIs. First, a targeted method with stable isotope dilution assay (SIDA) using labeled peptides as internal standards was applied. Additionally, four different approaches for relative quantitation were conducted, in detail, iTRAQ labeled and label free quantitation (LFQ) combined with data dependent acquisition (DDA) and data independent acquisition (DIA). Quantitation was performed manually (Skyline and MASCOT) and with different proteomics software tools (PLGS, MaxQuant, and PEAKS X Pro).

INSTRUMENT(S): Exactive Plus

ORGANISM(S): Triticum Durum Triticum Aestivum (wheat)

TISSUE(S): Endosperm

SUBMITTER: Sabrina Geisslitz  

LAB HEAD: Katharina Scherf

PROVIDER: PXD034458 | Pride | 2022-10-13

REPOSITORIES: Pride

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Dataset's files

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20211123_2021-004_TEP_T_nano_600ng_a.raw Raw
20211123_2021-004_TEP_T_nano_600ng_c.raw Raw
20211123_2021-004_TEP_T_nano_600ng_wdh_b.raw Raw
20211123_2021-005_TEP_T_nano_600ng_a.raw Raw
20211123_2021-005_TEP_T_nano_600ng_b.raw Raw
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Publications

Absolute and relative quantitation of amylase/trypsin-inhibitors by LC-MS/MS from wheat lines obtained by CRISPR-Cas9 and RNAi.

Geisslitz Sabrina S   Islam Shahidul S   Buck Lukas L   Grunwald-Gruber Clemens C   Sestili Francesco F   Camerlengo Francesco F   Masci Stefania S   D'Amico Stefano S  

Frontiers in plant science 20220829

Quantitation of wheat proteins is still a challenge, especially regarding amylase/trypsin-inhibitors (ATIs). A selection of ATIs was silenced in the common wheat cultivar Bobwhite and durum wheat cultivar Svevo by RNAi and gene editing, respectively, in order to reduce the amounts of ATIs. The controls and silenced lines were analyzed after digestion to peptides by LC-MS/MS with different approaches to evaluate changes in composition of ATIs. First, a targeted method with stable isotope dilution  ...[more]

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