Evidence map›Paper›PMID 41258465›Full record

ArticleScientific reports2025

Engineering a new tripartite split-ccGFP system from Corynactis californica for detecting protein-protein interactions.

Thomas M Groseclose, Erin Kober, Jennifer Zupancic, Claire K McLelland, Lexy A Lujan, Yuliya A Kunde, Sarah C Mozden, Nileena Velappan, Antonietta M Lillo, Emilia Solomon and 3 more

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In one paragraph

Article in Scientific reports, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.

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

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

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

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

13 authors.

Thomas M Groseclose *Bioscience Division, Los Alamos National Laboratory, Los Alamos, NM, 87545, USA.
Erin Kober *Bioscience Division, Los Alamos National Laboratory, Los Alamos, NM, 87545, USA.
Jennifer ZupancicBioscience Division, Los Alamos National Laboratory, Los Alamos, NM, 87545, USA.
Claire K McLellandBioscience Division, Los Alamos National Laboratory, Los Alamos, NM, 87545, USA.
Lexy A LujanBioscience Division, Los Alamos National Laboratory, Los Alamos, NM, 87545, USA.
Yuliya A KundeBioscience Division, Los Alamos National Laboratory, Los Alamos, NM, 87545, USA.
Sarah C MozdenBioscience Division, Los Alamos National Laboratory, Los Alamos, NM, 87545, USA.
Nileena VelappanBioscience Division, Los Alamos National Laboratory, Los Alamos, NM, 87545, USA.
Antonietta M LilloBioscience Division, Los Alamos National Laboratory, Los Alamos, NM, 87545, USA.
Emilia SolomonBioscience Division, Los Alamos National Laboratory, Los Alamos, NM, 87545, USA.
Jacob YoderBioscience Division, Los Alamos National Laboratory, Los Alamos, NM, 87545, USA.
Geoffrey S WaldoBioscience Division, Los Alamos National Laboratory, Los Alamos, NM, 87545, USA.
Hau B NguyenBioscience Division, Los Alamos National Laboratory, Los Alamos, NM, 87545, USA. hau@lanl.gov.

Funding

Defense Threat Reduction Agency 1308140096Los Alamos National Laboratory Directed Research and Development program 20220807PRD4
6 · The paper itself

Abstract

Protein-protein interactions (PPIs) are critical to a range of biological processes and, consequently, aberrant interactions are implicated in many disorders. The study of the complex networks of PPIs promises to elucidate undiscovered roles in cellular processes and the mechanisms of disease. To accomplish this, tools to effectively sense PPIs are necessary. Effective PPI sensors must rapidly detect interactions in real-time with high sensitivity without perturbing the proteins of interest (POIs) under study. Split fluorescent proteins have previously been used to successfully monitor PPIs, in part due to the small size of the tags. Here, we developed an optimized tripartite split GFP system based on Corynactis californica GFP (ccGFP) to detect PPIs in vitro. In this sensor system, ccGFP fragments ccGFP10 and ccGFP11 are tagged to two POIs. PPIs can then be detected via fluorescence by complementation to the third fragment, ccGFP1-9, which reconstitutes functional ccGFP. The optimized ccGFP system shows improved detection kinetics and pH and temperature stability compared to a previous system. We then validated the sensor by monitoring PPIs in two model systems: attractive/repulsive coiled-coils and rapamycin-inducible FRB/FKBP heterodimerization. Finally, we developed an anti-tripartite ccGFP single-chain variable fragment (scFv), which could enable versatile detection of identified protein-protein complexes.

Indexed as

Green Fluorescent ProteinsProtein Interaction MappingAnimalsProtein BindingGreen Fluorescent ProteinsAntibodyDirected evolutionGreen fluorescent protein (GFP)Protein detectionProtein engineeringProtein fragment complementationProtein-protein interactionsProtein taggingSingle-chain variable fragment (scFv)Split fluorescent protein

Identifiers

PMID41258465
PMCPMC12630887

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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 OpenQuestion graph.