Proteomics

Dataset Information

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Integrated Proteomics and Machine Learning Reveal AHNAK as a Discriminative Biomarker of Atrial Fibrillation in Heart Failure


ABSTRACT: Plasma proteins were digested with trypsin and the resulting peptides were analyzed by nanoLC–MS/MS on an EASY-nLC 1200 system coupled to an Orbitrap Exploris 480 mass spectrometer. Peptides were loaded onto a self-packed C18 reversed-phase column (25 cm × 100 μm i.d.) and separated at 500 nL/min using a linear gradient with solvent A (0.1% formic acid, 2% acetonitrile in water) and solvent B (0.1% formic acid, 90% acetonitrile in water): 4–20% B (0–68 min), 20–32% B (68–82 min), 32–80% B (82–86 min), and 80% B wash (86–90 min). The MS was operated with FAIMS using compensation voltages of −70 V and −45 V. Full MS scans were acquired at 60,000 resolution over m/z 400–1,200, and MS/MS scans at 30,000 resolution (first mass m/z 110). The top 15 precursors were selected for HCD (NCE 27%) with 30 s dynamic exclusion; AGC target was 75%, intensity threshold 10,000 ions/s, and maximum injection time 100 ms. TurboTMT was disabled for label-free quantification. Raw data were processed in Proteome Discoverer v2.4.1.15 and searched against the Homo_sapiens_9606_PR_20201214.fasta database (75,777 entries) with a reverse decoy strategy. Trypsin was specified with up to two missed cleavages; minimum peptide length was 6 aa. Carbamidomethyl (C) was set as a fixed modification; oxidation (M), protein N-terminal acetylation, methionine loss, and methionine loss + acetylation were set as variable modifications (≤3 variable modifications per peptide). Precursor and fragment mass tolerances were 10 ppm and 0.02 Da, respectively. FDR was controlled at <1% at the PSM, peptide, and protein levels. For label-free quantification, peptide intensities were normalized by peptide-wise mean centering across samples followed by within-sample median normalization, and protein abundances were summarized as the median of normalized peptide values per protein for downstream analyses.

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Blood Plasma

SUBMITTER: Zhenbang Gu  

LAB HEAD: Chen Liu

PROVIDER: PXD073684 | Pride | 2026-06-21

REPOSITORIES: Pride

Dataset's files

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Action DRS
XD058LQB1.zip Other
XD058LQB1_A2026.raw Raw
XD058LQB1_A2032.raw Raw
XD058LQB1_A2074.raw Raw
XD058LQB1_A2084.raw Raw
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