Evidence map›Paper›PMID 41668352›Full record

ArticleJournal of the American Society for Mass Spectrometry2026

On the Utility of Infrared Photoactivation for Native Top-Down and Complex-Down Orbitrap Mass Spectrometry of Soluble Proteoform Complexes.

Cynthia Nagy, Linda B Lieu, Christopher Mullen, Graeme C McAlister, Rafael D Melani, Joshua D Hinkle, Luca Fornelli

Abstract read
In one paragraph

Article in Journal of the American Society for Mass Spectrometry, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.

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0citing papers in PubMed
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1 · What the graph read from it

What it found

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The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

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Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

0 citing papers in PubMed.

No citing paper in PubMed yet.

4 · The record

Corrections and comments

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5 · Who and what money

Authors and funding

7 authors.

Cynthia NagySchool of Biological Sciences, University of Oklahoma, Norman, Oklahoma 73019, United States.
Linda B LieuDepartment of Chemistry and Biochemistry, University of Oklahoma, Norman, Oklahoma 73019, United States.
Christopher MullenThermo Fisher Scientific, San Jose, California 95134, United States.
Graeme C McAlisterThermo Fisher Scientific, San Jose, California 95134, United States.
Rafael D MelaniThermo Fisher Scientific, San Jose, California 95134, United States.
Joshua D HinkleThermo Fisher Scientific, San Jose, California 95134, United States.
Luca FornelliSchool of Biological Sciences, University of Oklahoma, Norman, Oklahoma 73019, United States.ORCID 0000-0001-6334-1046

Funding

A multi-level mass spectrometry pipeline for the analysis of whole proteoforms and their complexesR35GM147397 · NIGMS · UNIVERSITY OF OKLAHOMA · PI Luca Fornelli · 2022 to 2026
$1.9M
NIGMS NIH HHS R35 GM147397
6 · The paper itself

Abstract

Cellular functions arise from the coordinated action of proteoforms, which typically form multiproteoform complexes (MPCs), rather than functioning as isolated molecular entities. Deciphering the architecture and composition of MPCs is essential for linking proteoform diversity to biological function. Native top-down (nTD MS) and complex-down mass spectrometry (CxD MS) have emerged as powerful strategies to characterize MPCs, offering intact mass analysis as well as gas-phase sequencing either at the level of the complete assembly or its constituent proteoform subunits. Because the attainable sequence coverage is highly influenced by the ion activation technique, expanding activation strategies is key to improving proteoform characterization. To this end, we implemented infrared (IR) activation for the analysis of soluble MPCs─alcohol dehydrogenase (ADH; 147 kDa tetramer), enolase (96 kDa dimer), and pyruvate kinase (PK; 232 kDa tetramer). IR photons were used to induce infrared multiphoton dissociation (IRMPD) and to enhance electron-based fragmentation via activated-ion electron transfer dissociation (AI-ETD), and performance was benchmarked against higher-energy collisional dissociation (HCD). For ADH (∼36 kDa subunits), AI-ETD, HCD, and IRMPD returned similar sequence coverages in nTD MS experiments (36, 38, and 34%, respectively), with complementary cleavages resulting in a combined 48% coverage. As subunit mass increased, radical-driven fragmentation provided a clear advantage: for PK (∼57 kDa subunits), AI-ETD achieved 28% sequence coverage─approximately 15% higher than HCD or IRMPD. Together, these results highlight IR irradiation─both as a standalone dissociation modality and as a complement to electron-based activation─as a versatile strategy to enhance proteoform-level sequencing in native and complex-down MS workflows.

Indexed as

Mass SpectrometryMultiprotein ComplexesAlcohol DehydrogenaseInfrared RaysPhosphopyruvate HydratasePyruvate KinaseAlcohol DehydrogenaseMultiprotein ComplexesPhosphopyruvate HydratasePyruvate Kinasecomplex-downelectron transfer dissociationmultiproteoform complexesnative top-downOrbitrapphotoactivation

Identifiers

PMID41668352
PMCPMC12964546

What Socratic holds

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Registered trials

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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the Socratic graph.