MassIVE MSV000096467

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LC-MS data of trypsin-digested human serum albumin with and without acetyl coA.

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Description

To study the acetylation of human serum albumin (HSA) with acetyl coA, the LC-MS study was carried out. This is a part of ICMR, New Delhi, India, sanctioned project titled Pseudoesterase activity of albumin as a determinant for serum cholesterol in rabbits. Sample processing protocol For this 1 ml of 1 mg per ml solution of fatty acid free human albumin (SIGMA) in 10 mM Tris HCl buffer, pH 7.38 was incubated with 495, 20, and 8.04 micromolar of acetyl-CoA (prepared in MilliQ water) in separate aliquots for 2 hours at 37 degree C. Control albumin solution was added with MilliQ water and incubated under similar conditions. 100 microlitre of each aliquot was denatured by boiling with 10 mM dithiothreitol for 10 mins followed by the addition of Iodoacetamide (90 mM, SIGMA) and incubation for 30 mins in the dark at 37 degree C. These were then subjected to trypsin digestion (2 microgram, Promega, MS grade, prepared in acetic acid) and kept overnight at 37 degree C. The reaction was stopped by the addition of 1 percent trifluoroacetic acid (TCI, LCMS grade). The sample clean-up with C18 resins column was performed and the aliquots were then sent for data acquisition. Data Processing protocol The MaxQuant software was used for the analysis of data. Under group-specific parameters acetyl K, acetyl (N-term), and (acetyl protein N-term) were selected as variable modifications whereas the Carbamidomethyl (C) was considered a fixed modification. The label-free quantification was done and the albumin sequence file (accession P02768) was taken as a reference for understanding the analysis. For digestion, trypsin/P type was selected with all other parameters considered as default. The different raw files obtained are named as follows and uploaded. S1= HSA only S2= HSA + AcetylCoA (495 micromolar) S3= HAS+ AcetylCoA (20 micromolar) S4= HSA + AcetylCoA (8.04 micromolar) [doi:10.25345/C5804XX37] [dataset license: CC0 1.0 Universal (CC0 1.0)]

Keywords: Human Serum Albumin, Acetyl CoA, LC-MS, Trypsin digestion ; Pseudoesterase ; DatasetType:Proteomics

Contact

Principal Investigators:
(in alphabetical order) 		Dibyajyoti Banerjee, Postgraduate Institute of Medical Education and Research, Chandigarh, India, India
Submitting User: deepakk1812
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