PXD048758 is an
original dataset announced via ProteomeXchange.
Dataset Summary
Title | SKAP2 acts downstream of CD11b/CD18 and regulates neutrophil effector function |
Description | Background: The importance of CD11b/CD18 expression in neutrophil effector functions is well known. Beyond KINDLIN3 and TALIN1, which are involved in the induction of the high-affinity binding CD11b/CD18 conformation, the signaling pathways that orchestrate this response remain incompletely understood. Method: We performed an unbiased screening method for protein selection by biotin identification (BioID) and investigated the KINDLIN3 interactome. We used liquid chromatography with tandem mass spectrometry as a powerful analytical tool. Generation of NB4 CD18, KINDLIN3 or SKAP2 knock out neutrophils was achieved using CRISPR-Cas9 technology and the cells were examined for their effector function using flow cytometry, live cell imaging, microscopy, adhesion or antibody-dependent cellular cytotoxicity (ADCC). Results: Among the 325 proteins significantly enriched, we identified Src Kinase-Associated Phosphoprotein2 (SKAP2), a protein involved in actin polymerization and integrin-mediated outside-in signaling. CD18 immunoprecipitation in primary or NB4 neutrophils demonstrated the presence of SKAP2 in the CD11b/CD18 complex at steady state. Under this condition, adhesion to plastic, ICAM-1 or fibronectin was observed in the absence of SKAP2, which could be abrogated by blocking the actin-rearrangements with latrunculin B. Upon stimulation of SKAP2-deficient neutrophils, adhesion to fibronectin was enhanced whereas CD18 clustering was strongly reduced. This response corresponded with significantly impaired CD11b/CD18-dependent NADPH oxidase activity, phagocytosis and cytotoxicity against tumor cells. Conclusion: Our results suggest that SKAP2 has a dual role. It may restrict CD11b/CD18-mediated adhesion only under resting conditions, but its major contribution lies in the regulation of dynamic CD11b/CD18-mediated actin-rearrangements and clustering as required for cellular effector functions of human neutrophils. |
HostingRepository | PRIDE |
AnnounceDate | 2024-10-22 |
AnnouncementXML | Submission_2024-10-22_06:30:45.490.xml |
DigitalObjectIdentifier | |
ReviewLevel | Peer-reviewed dataset |
DatasetOrigin | Original dataset |
RepositorySupport | Unsupported dataset by repository |
PrimarySubmitter | Floris van Alphen |
SpeciesList | scientific name: Homo sapiens (Human); NCBI TaxID: 9606; |
ModificationList | iodoacetamide derivatized residue |
Instrument | Orbitrap Fusion |
Dataset History
Revision | Datetime | Status | ChangeLog Entry |
0 | 2024-01-22 03:58:52 | ID requested | |
1 | 2024-02-16 02:02:11 | announced | |
⏵ 2 | 2024-10-22 06:30:45 | announced | 2024-10-22: Updated project metadata. |
Publication List
Keyword List
submitter keyword: phagocytosis, Src Kinase Associated Phosphoprotein 2 (SKAP2), CD11b/CD18 integrin, filamentous actin, antibody-dependent cellular cytotoxicity (ADCC), adhesion,neutrophils |
Contact List
Hanke L. Matlung |
contact affiliation | Dept. of Molecular Hematology, Sanquin Research, Amsterdam, the Netherlands, and Landsteiner Laboratory, University of Amsterdam, Amsterdam, the Netherlands |
contact email | h.matlung@sanquin.nl |
lab head | |
Floris van Alphen |
contact affiliation | Sanquin Research |
contact email | F.vanalphen@sanquin.nl |
dataset submitter | |
Full Dataset Link List
Dataset FTP location
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PRIDE project URI |
Repository Record List
[ + ]
[ - ]
- PRIDE
- PXD048758
- Label: PRIDE project
- Name: SKAP2 acts downstream of CD11b/CD18 and regulates neutrophil effector function