Purified protein complexes were separated by SDS-PAGE and gel lanes divided into 3 sections (A, B and C). Samples prepared in triplicate (1, 2 and 3). A nanoElute (Bruker) was attached in line to a timsTOF Pro equipped with a CaptiveSpray Source (Bruker, Hamburg, Germany). Chromatography was conducted at 40C through a 25cm reversed-phase C18 column (PepSep) at a constant flow-rate of 0.5 uL/min. Mobile phase A was 98/2/0.1% LC/MS grade H2O/ACN/FA (v/v/v) and phase B was LC/MS grade ACN with 0.1% FA (v/v). During a 108 min method, peptides were separated by a 3-step linear gradient (5% to 30% B over 90 min, 30% to 35% B over 10 min, 35% to 95% B over 4 min) followed by a 4 min isocratic flush at 95% for 4 min before washing and a return to low organic conditions. Experiments were run as data-dependent acquisitions with ion mobility activated in PASEF mode. MS and MS/MS spectra were collected with m/z 100 to 1700 and ions with z = +1 were excluded.
[doi:10.25345/C5639K93B]
[dataset license: CC0 1.0 Universal (CC0 1.0)]
Keywords: membrane protein
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Principal Investigators: (in alphabetical order) |
Balyn Zaro, UCSF, United States |
| Submitting User: | ZaroLabUCSF |
Chen Z, Mondal A, Abderemane-Ali F, Jang S, Niranjan S, Montaño JL, Zaro BW, Minor DL.
EMC chaperone-CaV structure reveals an ion channel assembly intermediate.
Nature. 2023 Jul;619(7969):410-419. Epub 2023 May 17.
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