Related Experiment Video

Updated: Aug 11, 2025

Novel RNA-Binding Proteins Isolation by the RaPID Methodology
11:19

Novel RNA-Binding Proteins Isolation by the RaPID Methodology

Published on: September 30, 2016

9.1K

The small molecule inhibitor NAV-2729 has a complex target profile including multiple ADP-ribosylation factor

Eric M Rosenberg1, Xiaoying Jian1, Olivier Soubias2

  • 1Laboratory of Cellular and Molecular Biology, Center for Cancer Research, National Cancer Institute, Bethesda, Maryland, USA.

The Journal of Biological Chemistry
|February 9, 2023
PubMed

Insights

NAV-2729 inhibits cancer cell proliferation by targeting Arf GTPase regulators, not Arf6 directly. This compound acts as a model PH domain-binding inhibitor, revealing complexities in small molecule target identification.

Area of Science:

  • Molecular Biology
  • Cancer Research
  • Drug Discovery

Background:

  • ADP-ribosylation factor (Arf) GTPases and their regulators are crucial in cancer progression.
  • NAV-2729 was previously identified as an Arf6 inhibitor that reduced uveal melanoma progression.

Purpose of the Study:

  • To evaluate the antiproliferative effects of NAV-2729 on various cancer cell types.
  • To elucidate the precise molecular targets and mechanisms of NAV-2729 action.

Main Methods:

  • Cell proliferation assays across multiple cell lines.
  • Arf6 knockdown experiments.
  • Protein binding assays and cellular distribution studies.
  • Target identification screens.

Main Results:

  • NAV-2729 inhibited proliferation in diverse cell lines, independent of Arf6 expression levels.
  • Direct binding to Arf6 was not observed; instead, NAV-2729 targeted Arf exchange factors and GTPase-activating proteins.
  • NAV-2729 bound to the PH domain of ASAP1, altering its cellular distribution, but ASAP1 knockdown did not fully replicate NAV-2729's effects.
  • Extensive screens identified 48 additional potential NAV-2729 targets.

Conclusions:

  • NAV-2729's antiproliferative activity is not mediated by direct Arf6 inhibition.
  • NAV-2729 functions as a PH domain-binding inhibitor, with ASAP1 being one identified target.
  • The study highlights the challenges in pinpointing small molecule targets and establishes NAV-2729 as a valuable tool for studying PH domain interactions.
Keywords:
ADP-ribosylation factorASAP1GTPaseGTPase-activating proteinenzyme inhibitorguanine nucleotide exchange factor

More Related Videos

An Optimized Quantitative Pull-Down Analysis of RNA-Binding Proteins Using Short Biotinylated RNA07:55

An Optimized Quantitative Pull-Down Analysis of RNA-Binding Proteins Using Short Biotinylated RNA

Published on: February 17, 2023

3.9K
Global Identification of Co-Translational Interaction Networks by Selective Ribosome Profiling06:58

Global Identification of Co-Translational Interaction Networks by Selective Ribosome Profiling

Published on: October 7, 2021

2.5K

Related Concept Videos

Types of RNA01:23

Types of RNA

Overview
Three main types of RNA are involved in protein synthesis: messenger RNA (mRNA), transfer RNA (tRNA), and ribosomal RNA (rRNA). These RNAs perform diverse functions and can be broadly classified as protein-coding or non-coding RNA. Non-coding RNAs play important roles in the regulation of gene expression in response to developmental and environmental changes. Non-coding RNAs in prokaryotes can be manipulated to develop more effective antibacterial drugs for human or animal use.
RNA...
64.3K
Riboswitches01:56

Riboswitches

Riboswitches are non-coding mRNA domains that regulate the transcription and translation of downstream genes without the help of proteins. Riboswitches bind directly to a metabolite and can form unique stem-loop or hairpin structures in response to the amount of the metabolite present. They have two distinct regions – a metabolite-binding aptamer and an expression platform.
The aptamer has high specificity for a particular metabolite which allows riboswitches to specifically regulate...
8.2K
Transducer Mechanism: Enzyme-Linked Receptors01:27

Transducer Mechanism: Enzyme-Linked Receptors

Enzyme-linked receptors are cell-surface receptors acting as an enzyme or associating with an enzyme intracellularly. They make excellent drug targets. Drugs can bind to the extracellular ligand-binding domain or directly affect their enzymatic domain and alter their activity.
Major types that are helpful drug targets include:
2.6K
Experimental RNAi02:15

Experimental RNAi

RNA interference (RNAi) is a cellular mechanism that inhibits gene expression by suppressing its transcription or activating the RNA degradation process. The mechanism was discovered by Andrew Fire and Craig Mello in 1998 in plants. Today, it is observed in almost all eukaryotes, including protozoa, flies, nematodes, insects, parasites, and mammals. This precise cellular mechanism of gene silencing has been developed into a technique that provides an efficient way to identify and determine the...
6.2K
Ribosome Profiling02:24

Ribosome Profiling

Ribosome profiling or ribo-sequencing is a deep sequencing technique that produces a snapshot of active translation in a cell. It selectively sequences the mRNAs protected by ribosomes to get an insight into a cell’s translation landscape at any given point in time.
Applications of ribosome profiling
Ribosome profiling has many applications, including in vivo monitoring of translation inside a particular organ or tissue type and quantifying new protein synthesis levels.
The technique...
3.6K