PXD022299 is an
original dataset announced via ProteomeXchange.
Dataset Summary
Title | Improving the identification and coverage of plant transmembrane proteins in Medicago using bottom-up proteomics |
Description | Plant transmembrane proteins (TMPs) are essential for normal cellular homeostasis, nutrient exchange and responses to environmental cues. Commonly used bottom-up proteomic approaches fail to identify a broad coverage of peptide fragments derived from TMPs. Here, we used mass spectrometry (MS) to compare the effectiveness of two solubilisation and protein cleavage methods to identify shoot-derived TMPs from the legume Medicago. We compared a urea solubilisation, trypsin Lys-C (UR-TLC) cleavage method to a formic acid solubilisation, cyanogen bromide and trypsin Lys-C (FA-CTLC) cleavage method. We assessed the effectiveness of these methods by (i) comparing total protein identifications, (ii) determining how many TMPs were identified, and (iii) defining how many peptides incorporate all, or part, of transmembrane domains (TMD) sequences. The results show that the FA-CTLC method identified nine-fold more TMDs, and enriched more TMPs than the UR-TLC method. FA-CTLC identified more TMPs, particularly transporters, whereas, UR-TLC preferentially identified TMPs with one TMD, particularly signalling proteins. The results suggest that combining plant membrane purification techniques with both the FA-CTLC and UR-TLC methods will achieve a more complete identification and coverage of TMPs. |
HostingRepository | PRIDE |
AnnounceDate | 2020-12-02 |
AnnouncementXML | Submission_2021-01-06_06:25:42.xml |
DigitalObjectIdentifier | |
ReviewLevel | Peer-reviewed dataset |
DatasetOrigin | Original dataset |
RepositorySupport | Unsupported dataset by repository |
PrimarySubmitter | Han-Chung Lee |
SpeciesList | scientific name: Medicago tuberculata; NCBI TaxID: 1867346; |
ModificationList | deaminated residue; iodoacetamide derivatized residue |
Instrument | Q Exactive HF |
Dataset History
Revision | Datetime | Status | ChangeLog Entry |
0 | 2020-11-02 07:57:15 | ID requested | |
1 | 2020-12-01 23:32:09 | announced | |
⏵ 2 | 2021-01-06 06:25:43 | announced | 2021-01-06: Updated publication reference for PubMed record(s): 33391307. |
Publication List
Lee HC, Carroll A, Crossett B, Connolly A, Batarseh A, Djordjevic MA, Using Bottom-Up Proteomics. Front Plant Sci, 11():595726(2020) [pubmed] |
Keyword List
submitter keyword: TMD, TMP |
Contact List
Michael Djordjevic |
contact affiliation | ANU, RSB |
contact email | Michael.Djordjevic@anu.edu.au |
lab head | |
Han-Chung Lee |
contact affiliation | Research school of biology, The Australian National University |
contact email | Han.Lee@anu.edu.au |
dataset submitter | |
Full Dataset Link List
Dataset FTP location
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PRIDE project URI |
Repository Record List
[ + ]
[ - ]
- PRIDE
- PXD022299
- Label: PRIDE project
- Name: Improving the identification and coverage of plant transmembrane proteins in Medicago using bottom-up proteomics