This paper provides a detailed set of data on how the stal-lion sperm proteome differs among stallions with different sperm motilities, although within normal ranges. Findings distinguish proteins that may help to identify stallions of su-perior sperm motility. Sperm proteins were analyzed using a UHPLC/MS/MS system comprising of an Agilent 1290 in-finity series UHPLC coupled to an Agilent 6550 Q-TOF mass spectrometer (Agilent Technologies, Santa Clara, CA, USA). These data can be used to disclose potential targets to iden-tify good sperm samples and to study specific pathways in-volved in the regulation of sperm motility. This data article is linked to the paper "Proteins involved in mitochondrial metabolic functions and fertilization predominate in stallions with better motility Journal of Proteomics 247:104335 doi: 10.1016/j.jprot.2021.104335".(c) 2022 The Author(s). Published by Elsevier Inc.This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/

da Silva-Álvarez, Eva

Gaitskell-Phillips, Gemma

Gil, Maria C

Martín-Cano, Francisco E

Ortega-Ferrusola, Cristina

Ortiz-Rodríguez, José M

Peña, Fernando J

Silva-Rodríguez, Antonio

English

PubMed

Martín-Cano, Francisco E

Ortiz-Rodríguez, José M

Silva-Rodríguez, Antonio

da Silva-Álvarez, Eva

Peña, Fernando J

Archivio istituzionale della ricerca - Alma Mater Studiorum Università di Bologna

Data in Brief 45 (2022) 108578 Contents lists available at ScienceDirect Data in Brief journal homepage: www.elsevier.com/locate/dib Data Article Dataset of the sperm proteome of stallions with different motility Gemma Gaitskell-Phillips a , Francisco E. Martín-Cano a , José M Ortiz-Rodríguez a , Antonio Silva-Rodríguez b , Eva da Silva-Álvarez a , Maria C. Gil a , Cristina Ortega-Ferrusola a , Fernando J. Peña a , ∗a Laboratory of Equine Reproduction and Spermatology, Department of Animal Medicine, Veterinary Teaching Hospital, University of Extremadura, Cáceres, Spain b SIPA University of Extremadura, Cáceres, Spain a r t i c l e i n f o Article history: Received 25 September 2021 Revised 29 July 2022 Accepted 30 August 2022 Available online 5 September 2022 Dataset link: Stallion sperm proteins in 10 stallions with different motility (Original data) Keywords: Spermatozoa Proteomics Horses Motility CASA a b s t r a c t This paper provides a detailed set of data on how the stal- lion sperm proteome differs among stallions with different sperm motilities, although within normal ranges. Findings distinguish proteins that may help to identify stallions of su- perior sperm motility. Sperm proteins were analyzed using a UHPLC/MS/MS system comprising of an Agilent 1290 in- finity series UHPLC coupled to an Agilent 6550 Q-TOF mass spectrometer (Agilent Technologies, Santa Clara, CA, USA). These data can be used to disclose potential targets to iden- tify good sperm samples and to study specific pathways in- volved in the regulation of sperm motility. This data article is linked to the paper “Proteins involved in mitochondrial metabolic functions and fertilization predominate in stallions with better motility Journal of Proteomics 247:104335 doi: 10.1016/j.jprot.2021.104335”. © 2022 The Author(s). Published by Elsevier Inc. This is an open access article under the CC BY-NC-ND license ( http://creativecommons.org/licenses/by-nc-nd/4.0/ ) DOI of original article: 10.1016/j.jprot.2021.104335 ∗ Corresponding author. E-mail address: fjuanpvega@unex.es (F.J. Peña). Social media: @Fjpvega (F.J. Peña) https://doi.org/10.1016/j.dib.2022.108578 2352-3409/© 2022 The Author(s). Published by Elsevier Inc. This is an open access article under the CC BY-NC-ND license ( http://creativecommons.org/licenses/by-nc-nd/4.0/ ) 2 G. Gaitskell-Phillips, F.E. Martín-Cano and J.M. Ortiz-Rodríguez et al. / Data in Brief 45 (2022) 108578 SV  1 T  o  H  p  (  m  r  fi  t  t  a  p  t  pecifications Table Subject Veterinary Sciences; Reproductive Biology. Specific subject area Proteomics of stallion spermatozoa. Type of data Raw data How data were acquired Samples were analyzed using a UHPLC/MS/MS system consisting of an Agilent 1290 infinity series UHPLC coupled to an Agilent 6550 Q-TOF mass spectrometer (Agilent Technologies, Santa Clara, CA, USA) Data format Processed Analyzed Parameters for data collection Equine spermatozoa from 10 fertile stallions with a median age of 10.8 years with different values of motility estimated using computer assisted sperm analysis (CASA). Proteins were extracted from the spermatozoa of these stallions and analyzed using UHPLC/MS/MS. Description of data collection Comparative mass spectrometry proteomic profiling of the sperm proteome of fertile stallions with different motility. Data source location Cáceres, Extremadura, Spain. Data accessibility ProteomeXchange Consortium via the PRIDE partner repository with the dataset identifier PXD025807 ProteomeXchange title: Stallion sperm proteins in 10 stallions with different motility ProteomeXchange accession: PXD025807 PubMed ID: 34298182 ProjectWebpage: https://www.ebi.ac.uk/pride/archive/projects/PXD025807 FTP download: http://ftp.pride.ebi.ac.uk/pride/data/archive/2021/07/PXD025807/ Related research article Gaitskell-Phillips G, Martín Cano FE, Ortiz Rosdríguez JM, Silva Rodríguez A, Rojo-Domínguez P, Tapia JA, Gil MC, Ortega Ferrusola C, Peña FJ (2021). Proteins involved in metabolic functions and fertilization predominate in stallions with better motility Journal of Proteomics 247:104335 https://doi.org/10.1016/j.jprot.2021.104335 alue of the Data • The proteomic data reported here show that stallions with different sperm motility presentnotable differences in their proteome. • Re-analysis of the data set may help to characterize traits related to sperm motility in stal-lions and the influence of factors such as breed or age in the motility. . Data Description A data set of 903 proteins obtained from equine spermatozoa is provided (Supplementaryable 1). Proteins were extracted from spermatozoa from 10 fertile stallions, of a median agef 10.8 years old, of the following breeds; Arab, Andalusian, Andalusian-Arab, Spanish Sportorse, Lusitanian and Anglo-Arabian. Relative abundance of proteins is also provided. Theseroteins were obtained from 10 stallions with different motility and classified as having good > 90% total motility), average (between 80 and 90% total motility), and poor < 80% totalotility), although all of them have sperm parameters within normal ranges [1] . Motility pa-ameters were studied using Computer Assisted Sperm Analysis (CASA), including motility de-ned as the percentage of motile spermatozoa, linear motility, as the percentage of sperma-ozoa showing a linear pattern of movement, VCL defined as the actual velocity along therue trajectory in μm/s, VAP defined as the average velocity in μm/s, and the VSL defineds the straight-line velocity in μm/s. This information, after re-analysis, may be used to im-rove our understanding of the regulation of stallion sperm motility, and to develop on farmests to check sperm quality. The data set has been deposited to the ProteomeXchange Con-G. Gaitskell-Phillips, F.E. Martín-Cano and J.M. Ortiz-Rodríguez et al. / Data in Brief 45 (2022) 108578 3                                        sortium [2] via the PRIDE partner repository with the dataset identifier PRIDE PXD025807,https://www.ebi.ac.uk/pride/archive/projects/PXD025807 2. Experimental Design, Materials and Methods 2.1. Sample Preparation The same protocol as described in [3–9] was used. Stallion ejaculates were washed in PBS(600gx 10’), then the pellet formed by the spermatozoa was frozen immediately in liquid nitro-gen and kept frozen at -80 °C. Once the pellets were thawed the absence of debris and poten-tial contaminating cells was assessed using phase contrast microscopy. For the analysis, proteinswere extracted from 200 × 10 6 spermatozoa. The protein extraction was initiated with the sol-ubilization of the sperm proteins in the lysis buffer which composition follows, (C7:C7Bz0 [3-(4-heptyl) phenyl-(3-hydroxypropyl) dimethylammoniopropanesulfonate], 7M urea, 2M thiourea and 40 mM Tris (pH 10.4) as described in [1] . For the quantification of proteins we used the 2-DQuant Kit (GE Healthcare, Sevilla Spain) as indicated by the manufacturer, to normalize samplesto 100 μg of protein. In the next step 200 μL of the solution obtained containing 100 μg ofprotein was mixed with 100 μl of 25 mM ammonium bicarbonate buffer pH 8.5 (100 μg ofprotein in 300 μL of solution). Then, proteins were reduced by adding 30 μL of 10 mM DTTand incubated at 56 °C for 20 min. In the next step, proteins were alkylated by adding 30 μLof 20 mM IAA solution, and incubated at room temperature in the dark for 30 min. The laststep was digestion of the proteins, adding 2 μL of Trypsin Proteomics Grade (Sigma) (Trypsinsolution: 1 g/L in 1 mM HCl) and incubating for at least 3 h at 37 °C. After incubation, the reac-tion was stopped with 10 μL of 0.1% formic acid, and the peptide solution was filtered through0.2 μm (hydrophilic PTFE) into 2 mL dark glass vials. Finally, the protein samples were dried us-ing a nitrogen current with the vial in a heating block at 35 °C. The dried samples were dilutedin 20 μl of buffer A, (water/acetonitrile/formic acid (94.9:5:0.1)). 2.2. UHPLC-MS/MS Analysis The separation and analysis were conducted in an Agilent 1290 Infinity II Series UHPLC (Agi-lent Technologies, Santa Clara, CA, USA) equipped with an automated multisampler module anda High Speed Binary Pump, coupled to a Mass Spectrometer (Agilent 6550 Q-TOF, Agilent Tech-nologies, Santa Clara, CA, USA) using an Agilent Jet Stream Dual electrospray (AJS-Dual ESI) in-terface, controlled by the MassHunter Workstation Data Acquisition software (Agilent Technolo-gies, Rev. B.06.01), following previously published protocols [ 2 , 4 , 5 , 9 ]. The digested proteins wereinjected onto an Agilent Advance Bio Peptide Mapping HPLC column (2.7 μm, 150 × 2.1 mm,Agilent technologies), at 55 °C, at a flow rate of 0.4 ml/min. The gradient program was initiatedwith 2% of buffer B (water/acetonitrile/formic acid, 10:89.9:0.1) remaining in isocratic mode for5 min, after this time, linearly was increased up to 45 % B for 40 min, and up to 95 % B overadditional 15 min and finally remained constant for 5 min. After this initial 70 min run, 5 minfollowed, for the conditioning of the column for the next run using the initial condition. Themass spectrometer operated in positive mode. The nebulizer gas pressure was set to 35 psi, thedrying gas flow was set to 10 l/min at a temperature of 250 °C, and the sheath gas flow wasset to 12 l/min at 300 °C. The voltages of the capillary spray, fragmentor and octopole RF Vppwere, respectively, 3500 V, 340 V and 750 V. Profile data were acquired for both MS and MS/MSscans in extended dynamic range mode. MS and MS/MS mass ranges were 50–1700 m/z andscan rates were 8 spectra/sec for MS and 3 spectra/sec for MS/MS. Auto MS/MS mode was usedwith precursor selection by abundance and a maximum of 20 precursors selected per cycle. Aramped collision energy was used with a slope of 3.6 and an offset of -4.8. The same ion wasrejected after two consecutive scans. 4 G. Gaitskell-Phillips, F.E. Martín-Cano and J.M. Ortiz-Rodríguez et al. / Data in Brief 45 (2022) 108578 2 T  [  m  t  a  c  g  t  m  s  t  t  n3 c  UD t  D cC C  o  O  qA C  2  1  (.3. Data Processing Data were analyzed using the Spectrum Mill MS Proteomics Workbench (Rev B.04.01, Agilentechnologies, Santa Clara, CA, USA) as described in previous publications from our laboratory 5 , 6 ]. The raw data were extracted under default conditions: selection of non fixed or variableodifications; [MH] + 50–10,0 0 0 m/z; maximum precursor charge + 5; retention time and m/zolerance ± 60 s; minimum signal-to-noise MS (S/N) 25; finding 12 C signals. The MS/MS searchgainst the updated protein database (Uniprot/Horse) was done as follows: non fixed modifi-ations were selected and as variable modifications carbamidomethylated cysteines, tryptic di-estion with 5 maximum missed cleavages, ESI-Q-TOF instrument, minimum matched peak in-ensity 50%, maximum ambiguous precursor charge + 5, monoisotopic masses, peptide precursorass tolerance 20 ppm, product ion mass tolerance 50 ppm and calculation of reversed databasecores. The auto-validation strategy used was auto-threshold, in which the peptide score is au-omatically optimized for a target % FDR (1.2%), and immediately protein polishing validation ishen performed to increase the sequence coverage of validated results with the restriction of aew maximum target protein FDR (0 %). . Ethics Statement Stallions were maintained following specific institutional and European regulations for animalare (Law 6/2913 June 11th and European Directive 2010/63/EU). The ethical committee of theniversity approved this study REF IB-20 0 08. eclaration of Competing Interest The authors declare that they have no known competing financial interests or personal rela-ionships which have, or could be perceived to have, influenced the work reported in this article.ata Availability Stallion sperm proteins in 10 stallions with different motility (Original data) (ProteomeX-hange). RediT Author Statement Gemma Gaitskell-Phillips: Methodology, Writing – review & editing; Francisco E. Martín-ano: Methodology; José M Ortiz-Rodríguez: Methodology; Antonio Silva-Rodríguez: Method-logy; Eva da Silva-Álvarez: Methodology; Maria C. Gil: Supervision, Formal analysis; Cristinartega-Ferrusola: Supervision, Formal analysis; Fernando J. Peña: Formal analysis, Funding ac-uisition, Writing – review & editing, Supervision, Validation. cknowledgments The authors received financial support for this study from the Ministerio de Economía yompetitividad-FEDER, Madrid, Spain, grants, AGL2017- 83149- R Junta de Extremadura-FEDER (IB0 0 08 and GR 21060). JMOR holds a PhD grant from the Junta de Extremadura- FEDER (PD8005), Mérida, Spain, GG-P holds a PhD grant from the Ministry of Science, Madrid, SpainPRE2018-083354). G. Gaitskell-Phillips, F.E. Martín-Cano and J.M. Ortiz-Rodríguez et al. / Data in Brief 45 (2022) 108578 5                   Supplementary Materials Supplementary material associated with this article can be found, in the online version, atdoi: 10.1016/j.dib.2022.108578 . References [1] Gaitskell-Phillips G., Martín Cano F.E., Ortiz Rosdríguez J.M., Silva Rodríguez A., Rojo-Domínguez P., Tapia J.A., Gil M.C.,Ortega Ferrusola C., Peña F.J. (2021). Stallion sperm proteins in 10 stallions with different motility. https://www.ebi.ac.uk/pride/archive/projects/PXD025807 . [2] G. Gaitskell-Phillips , F.E. Martin-Cano , J.M. Ortiz-Rodriguez , A. Silva-Rodriguez , E. da Silva-Alvarez , P. Rojo-Dominguez ,et al. , Proteins involved in mitochondrial metabolic functions and fertilization predominate in stallions with bettermotility, J. Proteom. 247 (2021) 104335 . [3] Y. Perez-Riverol , J. Bai , C. Bandla , D. Garcia-Seisdedos , S. Hewapathirana , S. Kamatchinathan , et al. , The PRIDEdatabase resources in 2022: a hub for mass spectrometry-based proteomics evidences, Nucleic Acids Res. 50 (2022)D543–DD52 . [4] G. Gaitskell-Phillips , F.E. Martin-Cano , J.M. Ortiz-Rodriguez , A. Silva-Rodriguez , E. da Silva-Alvarez , M.C. Gil , et al. ,The seminal plasma proteins peptidyl arginine deaminase 2, rRNA adenine N (6)-methyltransferase and KIAA0825are linked to better motility post thaw in stallions, Theriogenology 177 (2022) 94–102 . [5] G. Gaitskell-Phillips , F.E. Martin-Cano , J.M. Ortiz-Rodriguez , E. da Silva-Alvarez , J. Masot , E. Redondo , et al. , Seminalplasma proteins as potential biomarkers for sperm motility and velocities, Theriogenology 177 (2022) 34–41 . [6] G Gaitskell-Phillips , FE Martin-Cano , JM Ortiz-Rodriguez , A Silva-Rodriguez , MC Gil , C Ortega-Ferrusola , et al. , In stal-lion spermatozoa, superoxide dismutase (Cu-Zn) (SOD1) and the aldo-keto-reductase family 1 member b (AKR1B1)are the proteins most significantly reduced by cryopreservation, J. Proteome Res. 20 (2021) 2435–2446 . [7] G. Gaitskell-Phillips , F.E. Martin-Cano , J.M. Ortiz-Rodriguez , A. Silva-Rodriguez , M.C. Gil , C. Ortega-Ferrusola , et al. ,Differences in the proteome of stallion spermatozoa explain stallion-to-stallion variability in sperm qualitypost-thaw, Biol. Reprod. 104 (2021) 1097–1113 . [8] G. Gaitskell-Phillips , F.E. Martin-Cano , J.M. Ortiz-Rodriguez , A. Silva-Rodriguez , H. Rodriguez-Martinez , M.C. Gil , et al. ,Seminal plasma AnnexinA2 protein is a relevant biomarker for stallions which require removal of seminal plasma forsperm survival upon refrigerationd, Biol. Reprod. 103 (2020) 1275–1288 . [9] F.E. Martin-Cano , G. Gaitskell-Phillips , J.M. Ortiz-Rodriguez , A. Silva-Rodriguez , A. Roman , P. Rojo-Dominguez , et al. ,Proteomic profiling of stallion spermatozoa suggests changes in sperm metabolism and compromised redox regula-tion after cryopreservation, J. Proteom. 221 (2020) 103765 . 

Dataset of the sperm proteome of stallions with different motility

This paper provides a detailed set of data on how the stallion sperm proteome differs among stallions with different sperm motilities, although within normal ranges. Findings distinguish proteins that may help to identify stallions of superior sperm motility. Sperm proteins were analyzed using a UHPLC/MS/MS system comprising of an Agilent 1290 infinity series UHPLC coupled to an Agilent 6550 Q-TOF mass spectrometer (Agilent Technologies, Santa Clara, CA, USA). These data can be used to disclose potential targets to identify good sperm samples and to study specific pathways involved in the regulation of sperm motility. This data article is linked to the paper “Proteins involved in mitochondrial metabolic functions and fertilization predominate in stallions with better motility Journal of Proteomics 247:104335 doi: 10.1016/j.jprot.2021.104335”

Martín-Cano, Francisco E.

Gil, Maria C.

Peña, Fernando J.

PubMed Central

http://dx.doi.org/10.1016/j.dib.2022.108578

Dataset of the sperm proteome of stallions with different motility

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