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LC-MS data of trypsin-digested human serum albumin with and without ester substrates (2-naphthyl acetate and p-nitrophenyl acetate) for different incubation times (45 minutes and 24 hours).

ABSTRACT: Project description To confirm the interaction or acetylation pattern of the human serum albumin (HSA) with ester substrates (2 naphthyl acetate and p nitrophenyl acetate), the LCMS 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.

INSTRUMENT(S): Orbitrap Fusion

ORGANISM(S): Homo Sapiens (ncbitaxon:9606)

SUBMITTER: Dibyajyoti Banerjee

PROVIDER: MSV000095594 | MassIVE | Tue Aug 13 23:22:00 BST 2024

REPOSITORIES: MassIVE

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Project description:This is study done for academic purpose and funded by ICMR, New Delhi, India, sanctioned project titled:- To study the acetylation of human serum albumin (HSA) in presence and absence of Glyphosate using 2-Naphthyl Acetate (2NA) as substrate and digestion with Pepsin. Project File No. 5/4/1-25/2020-NCD-1. Sample processing protocol: For this, 1 ml of 1 mg/ml solution of fatty acid free human albumin (SIGMA) in 10 mM Tris HCl buffer, pH 7.38 was incubated with Glyphosate (in Acetonitrile)(working concentration of 5 microMolar (Organophosphorous) for 1 hour at 37 degree Celsius. Further, acetylation was done using 2NA (prepared in Acetonitrile) for 45 minutes at 37 degree Celsius. 100 microlitre of each aliquot was denatured by boiling with 10 mM dithiothreitol for 10 minutes followed by the addition of Iodoacetamide (90 mM, SIGMA) and incubation for 30 mins in the dark at 37 degree Celsius. These were then subjected to Pepsin digestion (2 microgram, Promega, MS grade, prepared in acidic water).Next, samples to be digested by pepsin were incubated for 2 hours at 37 degree Celsius. Pepsin digestion was stopped by heating samples at 95 degree Celsius. 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 (2.0.3.1) 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, Pepsin enzyme was added to MaxQuant enzyme file. The albumin sequence file (accession: P02768) was taken as a reference for understanding the analysis. Description of the raw files is as follow. HSA_Glyphosate is Human serum albumin treated with Glyphosate and HSA_Glyphosate _2NA is Human serum Albumin treated with Glyphosate and then substrate (2NA) was added.

Project description:This is study done for academic purpose and funded by ICMR, New Delhi, India, sanctioned project titled:- To study the acetylation of human serum albumin (HSA) in presence and absence of Dichlorvos using 2-Naphthyl Acetate (2NA) as substrate and digestion with Pepsin. Project File No. 5/4/1-25/2020-NCD-1. Sample processing protocol: For this, 1 ml of 1 mg/ml solution of fatty acid free human albumin (SIGMA) in 10 mM Tris HCl buffer, pH 7.38 was incubated with Dichlorvos (in Acetonitrile)(working concentration of 5 microMolar (Organophosphorous) for 1 hour at 37 degree Celsius. Further, acetylation was done using 2NA (prepared in Acetonitrile) for 45 minutes at 37 degree Celsius. 100 microlitre of each aliquot was denatured by boiling with 10 mM dithiothreitol for 10 minutes followed by the addition of Iodoacetamide (90 mM, SIGMA) and incubation for 30 mins in the dark at 37 degree Celsius. These were then subjected to Pepsin digestion (2 microgram, Promega, MS grade, prepared in acidic water).Next, samples to be digested by pepsin were incubated for 2 hours at 37 degree Celsius. Pepsin digestion was stopped by heating samples at 95 degree Celsius. 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 (2.0.3.1) 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, Pepsin enzyme was added to MaxQuant enzyme file. The albumin sequence file (accession: P02768) was taken as a reference for understanding the analysis. Description of the raw files is as follow. HSA_Dichlorvos is Human serum albumin treated with Dichlorvos and HSA_Dichlorvos _2NA is Human serum Albumin treated with Dichlorvos and then substrate (2NA) was added.

Dataset Information

LC-MS data of trypsin-digested human serum albumin with and without ester substrates (2-naphthyl acetate and p-nitrophenyl acetate) for different incubation times (45 minutes and 24 hours).

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