PXD037181-1

PXD037181 is an original dataset announced via ProteomeXchange.

Title	Characterization of acrosin and acrosin binding protein as novel CRISP2 interacting proteins in boar spermatozoa
Description	Previously we have reported that cysteine rich secretory protein 2 (CRISP2) is involved in high molecular weight complexes in boar spermatozoa under a native state. In the current study, CRISP2 interacting partners were identified using proteomic technology and their interaction was investigated under non-capacitating conditions, after the induction of in vitro sperm capacitation and then subsequently during an ionophore A23187 induced acrosome reaction. Blue native gel electrophoresis and native immunoblots revealed that CRISP2 was present within a ~150 kDa protein complex under all three conditions. Interrogation of CRISP2-immunoreactive bands from blue native gels as well as CRISP2 immunoprecipitated products using mass spectrometry consistently revealed that acrosin and acrosin binding protein (ACRBP) were among the most abundant proteins and present under the three conditions. Co-immunoprecipitation and immunoblotting indicated that CRISP2 interacted with pro-acrosin (~53 kDa) and ACRBP under all three conditions and additionally to acrosin (~35 kDa) after capacitation and the acrosome reaction. The colocalization of these interacting proteins with CRISP2 was assessed via in situ proximity ligation assays. The colocalization signal of CRISP2 and acrosin in the acrosome seemed dispersed after capacitation but was consistently present in the sperm tail under all conditions. The fluorescent foci of CRISP2 and ACRBP colocalization appeared to be redistributed within the sperm head from the anterior acrosome to the post acrosomal sheath region upon capacitation. These results suggest that CRISP2 may act as a scaffold for protein complexes formation to ensure the correct positioning of proteins required for the acrosome reaction and zona pellucida penetration.
HostingRepository	PRIDE
AnnounceDate	2023-03-09
AnnouncementXML	Submission_2023-03-09_04:47:54.234.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	RiccardoZenezini Chiozzi
SpeciesList	scientific name: Sus scrofa domesticus (domestic pig); NCBI TaxID: 9825;
ModificationList	phosphorylated residue
Instrument	Q Exactive

Revision	Datetime	Status	ChangeLog Entry
0	2022-10-05 00:49:01	ID requested
⏵ 1	2023-03-09 04:47:54	announced
2	2023-11-14 08:58:51	announced	2023-11-14: Updated project metadata.

submitter keyword: boar spermatozoa

protein complexes

colocalization

capacitation, interacting proteins

Albert JRHeck
contact affiliation	Biomolecular Mass Spectrometry and Proteomics, Bijvoet Centre for Biomolecular Research and Utrecht Institute for Pharmaceutical Sciences, Utrecht University, The Netherlands
contact email	a.j.r.heck@uu.nl
lab head
RiccardoZenezini Chiozzi
contact affiliation	UCL Mass Spectrometry Science Technology Platform, Division of Biosciences, University College London, London
contact email	r.zenezinichiozzi@uu.nl
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD037181
     1. Label: PRIDE project
     2. Name: Characterization of acrosin and acrosin binding protein as novel CRISP2 interacting proteins in boar spermatozoa