PXD071784-1

PXD071784 is an original dataset announced via ProteomeXchange.

Title	Parallel Reaction Monitoring to Assess High-Affinity N-Glycoforms of the Human Fcγ Receptor IIIa (CD16a)
Description	Fc γ-receptor IIIa (FcγRIIIa, or CD16a) is a receptor for the Fc region of IgG antibodies and plays an essential role in the regulation of antibody effector functions. While the activation of CD16a plays a role in a healthy immune response, dysregulated CD16a activity may contribute to tissue damage and chronic inflammation by over-activating immune cells or, conversely, by compromising the removal of immune complexes. Functionally distinct glycosylated forms of CD16a exist, with CD16a glycoforms modified with high mannose N-linked glycans at site N162 demonstrating a higher binding affinity for IgG compared to other forms of CD16a. Here, we developed and tested a nano-liquid chromatography tandem mass spectrometry (nLC-MS/MS) parallel reaction monitoring (PRM) method for the site-specific study of CD16a glycosylation. The PRM method was tested by spiking recombinant CD16a (F158 or V158) into a complex matrix consisting of THP-1 cell lysate digest, then the CD16a glycopeptide PRM method was applied to measure statistically significant changes in site N162 N-glycan compositions corresponding to high affinity glycoforms of CD16a in a model of monocyte differentiation. Analyses were extended to the analysis of CD16a derived from human primary monocytes and peripheral blood mononuclear cells. CD16a site N162 N-glycosylation demonstrated remarkable similarity across the surveyed cell types, including in THP-1 cells – a monocyte cell line isolated from the peripheral blood of an acute monocytic leukemia patient, as well as primary human monocytes, and PBMCs. Critically, CD16a glycopeptide PRM facilitated measurement of changes in CD16a glycosylation at site N162, with high mannose N-glycan compositions, corresponding to high affinity glycoforms of CD16a, increasing in a model of monocyte differentiation into macrophages. Here, we have demonstrated the feasibility of utilizing a glycopeptide nLC-MS/MS-PRM targeted approach to study CD16a glycosylation in material from primary cells in a glycosylation site-specific manner. This strategy is well-suited for adaptation to clinical studies focused on understanding the role of CD16a glycosylation in physiology and disease, with a focus on immune disorders.
HostingRepository	PRIDE
AnnounceDate	2026-04-27
AnnouncementXML	Submission_2026-04-26_19:12:58.793.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Carlos Pavan
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: NEWT:9606;
ModificationList	iodoacetamide derivatized residue
Instrument	Orbitrap Eclipse

Revision	Datetime	Status	ChangeLog Entry
0	2025-12-09 15:26:12	ID requested
⏵ 1	2026-04-26 19:12:59	announced

10.1021/acs.jproteome.5c01234;

Pavan CH, Desai J, Medina Navarro V, Chandler KB, Receptor IIIa (CD16a) via Parallel Reaction Monitoring Mass Spectrometry. J Proteome Res, 25(6):2877-2890(2026) [pubmed]

submitter keyword: glycosylation, CD16a, oligomannose, IgG, N162, immunotherapy, high mannose N-linked glycans,FcγRIIIa, parallel reaction monitoring, monocytes, Fc γ (gamma) receptor IIIa

Kevin Brown Chandler
contact affiliation	Assistant Professor Department of Cellular and Molecular Medicine Florida International University | Herbert Wertheim College of Medicine
contact email	kchandle@fiu.edu
lab head
Carlos Pavan
contact affiliation	Florida International University
contact email	cpavan@fiu.edu
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD071784
     1. Label: PRIDE project
     2. Name: Parallel Reaction Monitoring to Assess High-Affinity N-Glycoforms of the Human Fcγ Receptor IIIa (CD16a)