Use of an Intramolecular Quenched Fluorescence (IQF) Cleavage Assay for Assessing Enzyme Kinetics of Gamma-Secretase in Human Skin Fibroblasts and Keratinocytes.

Beita Badiei, Luis A Garza

Abstract read

In one paragraph

Article in Methods in molecular biology (Clifton, N.J.), 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

0cells of the map it votes in

0citing papers in PubMed

–field-weighted citation impact

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

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

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

2 authors.

Beita BadieiDepartment of Dermatology, Johns Hopkins University School of Medicine, Baltimore, MD, USA.

Luis A GarzaDepartment of Dermatology, Johns Hopkins University School of Medicine, Baltimore, MD, USA. LAG@jhmi.edu.

Funding

Inhibition of regeneration restraining pathways to promote healingR01AR083822 · NIAMS · JOHNS HOPKINS UNIVERSITY · PI Luis Andres Garza · 2025 to 2026

$1.3M

NIAMS NIH HHS R01 AR083822

6 · The paper itself

Abstract

This study describes an intramolecular quenching assay to evaluate gamma-secretase (GS) enzyme activity in human dermal cells. The method utilizes a fluorogenic peptide substrate, mimicking a fragment of amyloid precursor protein (APP), in which a quencher suppresses the fluorescence of a fluorophore until enzymatic cleavage occurs, resulting in a measurable increase in fluorescence. This real-time, direct measurement of GS activity allows for precise kinetic analysis using Michaelis-Menten modeling to define Kd and Vmax. The assay is designed to quantify GS activity in human dermal fibroblasts and keratinocytes, enabling comparison between samples derived from hidradenitis suppurativa (HS) patients and healthy controls, as well as investigating the effects of subunit knockdown, such as nicastrin, on GS function. The method offers several advantages, including simplicity, cost-effectiveness, and adaptability for high-throughput screening for GS enzyme inhibitors.

Indexed as

Amyloid precursor proteinDermal fibroblastsEnzyme kineticsFluorescenceGamma-secretaseHidradenitis suppurativaKeratinocytesMichaelis–MentenNicastrinPeptide cleavage

Identifiers

PMID39776075

DOI10.1007/7651_2024_587

PMCPMC12392851

What OpenQuestion holds

Textmetadata

LicenceTDM

Read underepoch 390

Registered trials

None linked

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.