MassIVE MSV000094146

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Disrupted Nitric Oxide Homeostasis Impacts Fertility Through Multiple Processes Including Protein Quality Control

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Description

Pooled sepals and petals (hereafter referred to as SePel) of hot5-2 and WT were harvested from the same plants of which pistils have been isolated. Material from around 50 flowers at stages before pollination (floral developmental stage, FD 12-13) were pooled and represent one biological replicate. A total of five biological replicates were further processed as mentioned for the pistil proteome. For the LC-MS\MS run, a 3 uL injection was loaded by a Thermo Easy nLC 1000 UPLC on to a 2 cm trapping column and desalted with 8 uL mobile phase A (0.1% formic acid in water). Peptides were eluted at 300 nL/min on to a 75 um x 15 cm RSLC column (Thermo) using a linear gradient of 5-35% mobile phase B (0.1% formic acid in acetonitrile) over 90 minutes. Ions were introduced by positive ESI using a stainless steel capillary at 2.1 kV into a Thermo Orbitrap Fusion tribrid mass spectrometer. Mass spectra were acquired over m/z 300-1750 at 120,000 resolution (m/z 200) with an AGC target of 1e6, and data-dependent acquisition selected the top 10 most abundant precursor ions for tandem mass spectrometry by HCD fragmentation using an isolation width of 1.6 Da, maximum fill time 110 ms and AGC target 1e5. Peptides were fragmented with a normalized collision energy 27, and fragment spectra acquired at a resolution of 15,000 (m/z 200). [doi:10.25345/C5J67973H] [dataset license: CC0 1.0 Universal (CC0 1.0)]

Keywords: gsnor/hot5_2 quantitative SePel proteome, AT5G43940

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Principal Investigators:
(in alphabetical order) 		Elizabeth Vierling, University of Massachusetts Amherst, USA
Submitting User: PatrickTreffon
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