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PXD023929

PXD023929 is an original dataset announced via ProteomeXchange.

Dataset Summary
TitleMature proteins of LAG-3 with or without mutations were digested with chymotrypsin followed by data-dependent LC-MS/MS analysis
DescriptionThe FLAG-tagged soluble monomeric proteins of LAG-3-WT, LAG-3-V20A, and LAG-3-L14Q/V20A were immunoprecipitated from culture supernatants of Plat-E cells transfected with the corresponding plasmid vectors using ANTI-FLAG M2 Magnetic Beads. Proteins on the beads were digested with chymotrypsin (Roche) followed by reduction and alkylation. The peptides were subjected to data-dependent LC-MS/MS analysis using an EASY-nLC 1200 UHPLC connected to a Q Exactive Plus mass spectrometer through a nanoelectrospray ion source (Thermo Fisher Scientific). Raw data were directly analyzed against the SwissProt database restricted to H. sapiens supplemented with predicted amino acid sequences of LAG-3 with or without mutations starting from A18 to L29 using Proteome Discoverer version 2.3 (Thermo Fisher Scientific) with Sequest HT search engine. The search parameters were as follows: (a) chymotrypsin as an enzyme with up to two missed cleavages; (b) precursor mass tolerance of 10 ppm; (c) fragment mass tolerance of 0.02 Da; (d) carbamidomethylation of cysteine as a fixed modification; and (e) acetylation of protein N-terminus and oxidation of methionine as variable modifications. Peptides were filtered at a false discovery rate of 1% using the percolator node.
HostingRepositoryjPOST
AnnounceDate2022-04-11
AnnouncementXMLSubmission_2022-09-18_03:35:55.243.xml
DigitalObjectIdentifier
ReviewLevelPeer-reviewed dataset
DatasetOriginOriginal dataset
RepositorySupportUnsupported dataset by repository
PrimarySubmitterHidetaka Kosako
SpeciesList scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationListS-carboxamidomethyl-L-cysteine; alpha-amino acetylated residue; L-methionine sulfoxide
Instrumentinstrument
Dataset History
RevisionDatetimeStatusChangeLog Entry
02021-02-02 01:13:30ID requested
12022-04-10 08:00:04announced
22022-04-19 23:00:46announced2022-04-20: Updated PubMed.
32022-09-18 03:35:56announced2022-09-18: Updated FTP location.
Publication List
Maruhashi T, Sugiura D, Okazaki IM, Shimizu K, Maeda TK, Ikubo J, Yoshikawa H, Maenaka K, Ishimaru N, Kosako H, Takemoto T, Okazaki T, cell function and suppresses autoimmunity and anti-cancer immunity. Immunity, 55(5):912-924.e8(2022) [pubmed]
Keyword List
submitter keyword: DDA, mature LAG-3, N-terminal peptides
Contact List
Hidetaka Kosako
lab head
Hidetaka Kosako
contact affiliationTokushima University
dataset submitter
Full Dataset Link List
jPOST dataset URI
Dataset FTP location
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