Qualitative and Quantitative Analysis of Proteome ... - ACS Publications

Jun 28, 2017 - Intercollegiate Faculty of Biotechnology, University of Gdańsk and Medical University of Gdańsk, Abrahama 58, 80-307 Gdańsk,. Gdańs...
0 downloads 0 Views 2MB Size
Subscriber access provided by NEW YORK UNIV

Article

Qualitative and quantitative analysis of proteome and peptidome of human follicular fluid using multiple samples from single donor with LC-MS and SWATH methodology Aleksandra Ewa Lewandowska, Katarzyna Macur, Paulina Czaplewska, Joanna Liss, Krzysztof #ukaszuk, and Stanislaw Oldziej J. Proteome Res., Just Accepted Manuscript • DOI: 10.1021/acs.jproteome.7b00366 • Publication Date (Web): 28 Jun 2017 Downloaded from http://pubs.acs.org on June 29, 2017

Just Accepted “Just Accepted” manuscripts have been peer-reviewed and accepted for publication. They are posted online prior to technical editing, formatting for publication and author proofing. The American Chemical Society provides “Just Accepted” as a free service to the research community to expedite the dissemination of scientific material as soon as possible after acceptance. “Just Accepted” manuscripts appear in full in PDF format accompanied by an HTML abstract. “Just Accepted” manuscripts have been fully peer reviewed, but should not be considered the official version of record. They are accessible to all readers and citable by the Digital Object Identifier (DOI®). “Just Accepted” is an optional service offered to authors. Therefore, the “Just Accepted” Web site may not include all articles that will be published in the journal. After a manuscript is technically edited and formatted, it will be removed from the “Just Accepted” Web site and published as an ASAP article. Note that technical editing may introduce minor changes to the manuscript text and/or graphics which could affect content, and all legal disclaimers and ethical guidelines that apply to the journal pertain. ACS cannot be held responsible for errors or consequences arising from the use of information contained in these “Just Accepted” manuscripts.

Journal of Proteome Research is published by the American Chemical Society. 1155 Sixteenth Street N.W., Washington, DC 20036 Published by American Chemical Society. Copyright © American Chemical Society. However, no copyright claim is made to original U.S. Government works, or works produced by employees of any Commonwealth realm Crown government in the course of their duties.

Page 1 of 72

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60

Journal of Proteome Research

Qualitative and quantitative analysis of proteome and peptidome of human follicular fluid using multiple samples from single donor with LC-MS and SWATH methodology Aleksandra E. Lewandowska*,†, Katarzyna Macur†, Paulina Czaplewska†, Joanna Liss‡, Krzysztof Łukaszuk‡,§, Stanisław Ołdziej*,† †Intercollegiate Faculty of Biotechnology UG&MUG, University of Gdańsk, Abrahama 58, 80307 Gdańsk, Gdańsk, Poland. ‡INVICTA Fertility and Reproductive Center , Trzy Lipy 3, 80-172 Gdańsk, Gdańsk , Poland. §Department of Obstetrics and Gynecological Nursing, Faculty of Health Sciences, Medical University of Gdańsk, Dębinki 7, 80-211 Gdańsk, Gdańsk, Poland. Keywords: LC-MS, SWATH, human follicular fluid, proteome, peptidome, oocyte quality control

ACS Paragon Plus Environment

1

Journal of Proteome Research

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60

Page 2 of 72

ABSTRACT: Human follicular fluid (hFF) is a natural environment of oocyte maturation, and some components of hFF could be used to judge oocyte capability for fertilization and further development. In our pilot small scale study three samples from four donors (12 samples in total) were analyzed to determine which hFF proteins/peptides could be used to differentiate individual oocytes and which are patient specific. Ultrafiltration was used to fractionate hFF to High Molecular Weight (HMW) – proteome (>10kDa) and Low Molecular Weight (LMW) – peptidome (10 kDa) containing whole proteins, and low molecular weight fraction (LMW, 0.05 (10%). All identified hits from database search were manually inspected for presence of amino acids substitutions or modifications to lower the chances of possible false protein identification. Runs of LMW fraction samples were processed accordingly with changes in parameters: no modification of cysteines, and no enzyme specified in digestion. Only the protein identifications with 1% FDR in HMW fraction and peptide identifications with 1% FDR in LMW fraction were considered valid. Quantitative analysis was performed by loading the resulting from the ProteinPilot database search file to the MS/MS ALL with SWATH™ Acquisition MicroApp 2.01 in PeakView® 2.2 (SCIEX) to create automatically a spectral library file. The assumptions employed during the spectral library creation included: a maximum of 10 peptides per protein and 10 transitions per peptide; peptide modification was allowed (but shared peptides were excluded); only peptides of at least 95% confidence and an extraction window of 5 min and 0.02 Da width were used. The peptides profiles and transitions were manually inspected to ensure that PeakView extraction was accurate and that ion transitions correlated with each other. Proteins have been quantified using at least two peptides with a 95% identification certainty. The created spectral libraries were used for targeted data extraction from SWATH-MS experiments and further processing in MS/MS ALL with SWATH™ Acquisition MicroApp 2.01 in PeakView® 2.2 software. Statistical analysis of processed data was performed in MarkerView 1.2.1 Software (SCIEX). For each hFF sample three biological and three technical replicates were analyzed and the areas under XICs for technical replicates were averaged. Quantitative data was normalized using Total Area Sums

ACS Paragon Plus Environment

14

Page 15 of 72

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60

Journal of Proteome Research

(TAS) approach. The coefficient of variation (CV) was calculated using SWATH data from parallel measurements of four pool reference samples to monitor measurement variations for each protein (in HMWF) and peptide (in LMWF). The t-tests were performed twice, between groups composed of all samples for each individual oocyte, and groups composed of all samples from each patient, and the peptides/proteins with p