PXD025947

PXD025947 is an original dataset announced via ProteomeXchange.

Title	Quantification of oxidative modification of human serum albumin by MRM-HR
Description	Two sampling areas were focused in this study; Tande tande hamlet as high-level natural background radiation area (HBRA) and Topoyo sub-district as normal level background radiation area (NBRA), which was done under coordination with Local Health Office. The former located in 2° 44’ 38.3’’ south latitude/ 118° 48’ 47’’ east longitude belongs to Northern Botteng village in Simboro district of Mamuju city, and the population of the hamlet is 350 people in 2017 on an area of 16.22 km2 according to the information of BPS-Statistics of Mamuju Regency 2018. The latter located in 1° 59’ 38.3’’ south latitude/ 119° 28’ 16.7’’ east longitude belongs to Topoyo sub-district of Mamuju Tengah city about 120 km from Mamuju city, and the population of the sub-district is 31,888 people with 16,463 men (51.63%) and 15,425 women (48.37%) in 2017 on an area of 844.80 km2 according to the information of BPS-Statistics of Mamuju Regency 2019. 26 healthy adult inhabitants (13 males and 13 females) from Tande tande hamlet and 23 healthy adult inhabitants (14 males and 9 females) from Topoyo sub-district were included in this study. All volunteers were informed about the nature, aims, and intention of the study and signed a consent form and questionnaire to obtain information on age and occupation and another biodata before providing blood samples. Any individuals suffering from an illness or taking medication were excluded. Peripheral blood samples then were obtained from the antecubital vein using BD Vacutainer tubes in the morning (about from 6 a.m. to 9 a.m.) and placed at room temperature for at least 30 min to allow blood-clotting. Sera was collected by centrifugation at 1,200 ×g for 10 min at 4°C and stored at - 80°C until the analysis. 2 μl of sera was diluted with 198 μl of 50 mM ammonium bicarbonate. The serum proteins were precipitated by adding 1 ml of acetone. The precipitated proteins were collected by centrifugation at 15,000 ×g for 10 min. The precipitate was resuspended in 10 μl of 500 mM ammonium bicarbonate and denatured with an equivalent volume of trifluoroethanol and 5 μl of 200 mM DTT. The samples were incubated at 90°C for 30 min and cooled to room temperature. Free cysteine residues were alkylated with 10 μl of 200 mM iodoacetamide for 60 min at room temperature in the dark and the remaining iodoacetamide was quenched by adding 5 μl of 200 mM DTT. The samples were then mixed with 320 μl of 100 mM ammonium bicarbonate. The samples were incubated with 5 μg trypsin (TPCK treated, AB Sciex) at 37°C for 18 h. The samples were desalted with C18 ZipTip (Millipore, Bedford, MA, USA) and eluted with H2O/acetonitrile (5/5; v/v). The ZipTip eluates were dried in a vacuum centrifuge. Desalted samples were rehydrated in 0.1% formic acid and were analyzed by LC-MS using a nanoLC Eksigent 400 system (Eksigent, AB Sciex), coupled online to a TripleTOF6600 mass spectrometer (AB Sciex). Peptide separation was performed using liquid chromatography on a trap and elution configuration using a nano trap column (350 μm × 0.5 mm, 3 μm, 120 Å, AB Sciex) and a nano ChromXP C18 reverse-phase column (75 μm × 15 cm, 3 μm, 120 Å, AB Sciex) at 300 nl/min with a 90 min linear gradient of 8% to 30% acetonitrile in 0.1% formic acid, and then, with a 10 min linear gradient of 30% to 40% acetonitrile in 0.1% formic acid. On the basis of the result of the information-dependent acquisition, an assay for a high-resolution multiple reaction monitoring (MRM-HR) experiment targeted to the serum albumin oxidative modification was developed using Skyline software (MacCoss Lab, University of Washington, WA). The transitions of MRM-HR targeting each peptide were shown in Table 3. A TOF-MS scan in the mass range of 400-2,250 Da with 0.1 sec accumulation time was acquired followed by the product ion of target peptide precursors mass with 100 ms accumulation time. Collision energies were calculated with the rolling collision energy as in the information-dependent acquisition experiment. For MRM-HR data analysis, MultiQuant 3.0 Software (AB Sciex) was used to calculate integrated area of each peptide. All peak selections were checked manually after the automated matches.
HostingRepository	jPOST
AnnounceDate	2022-05-12
AnnouncementXML	Submission_2022-09-18_02:33:49.967.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Yota Tatara
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	S-carboxamidomethyl-L-cysteine; N6-acetyl-L-lysine; alpha-amino acetylated residue; 2-pyrrolidone-5-carboxylic acid (Gln); 2-pyrrolidone-5-carboxylic acid (Glu); L-methionine sulfoxide; phosphorylated L-histidine; O-phospho-L-serine; O-phospho-L-threonine; O4'-phospho-L-tyrosine
Instrument	TripleTOF 6600

Revision	Datetime	Status	ChangeLog Entry
0	2021-05-11 19:03:57	ID requested
1	2022-05-11 08:00:09	announced
2	2022-05-30 01:11:27	announced	2022-05-30: Updated PubMed.
⏵ 3	2022-09-18 02:33:50	announced	2022-09-18: Updated FTP location.

Yamaguchi M, Tatara Y, Nugraha ED, Ramadhani D, Tamakuma Y, Sato Y, Miura T, Hosoda M, Yoshinaga S, Syaifudin M, Kashiwakura I, Tokonami S, Detection of biological responses to low-dose radiation in humans. Free Radic Biol Med, 184():196-207(2022) [pubmed]

submitter keyword: human, serum, MRM-HR

Ikuo Kashiwakura
lab head
Yota Tatara
contact affiliation	Hirosaki University
dataset submitter

jPOST dataset URI

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