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Cited by 1Protein Kinase Regulation and GTPase SignalingCellular transport and secretionReceptor Mechanisms and Signaling

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An active allosteric mechanism in ASAP1-mediated Arf1 GTP hydrolysis redefines PH domain function

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Soubias, Olivier, Foley, Samuel L., Jian, Xiaoying, Jackson, Rebekah A., Zhang, Yue, Rosenberg, Eric M., Hu, Benjamin J., Li, Jess, Heinrich, Frank, Johnson, Margaret E., Sodt, Alexander J., Randazzo, Paul A., Byrd, R. Andrew·2025·Nature Communications
DOI 10.1038/s41467-025-63764-wPMC PMC12485133

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Abstract

GTPase-activating proteins are important regulators of small GTPases; among these, ASAP1 stimulates GTP hydrolysis on Arf1 and is implicated in cancer progression. ASAP1 contains a Pleckstrin Homology (PH) domain essential for maximum Arf·GTP hydrolysis. The prevailing view of PH domains is that they regulate proteins through passive mechanisms like membrane recruitment. In sharp contrast, we show that the PH domain of ASAP1 actively contributes to Arf1 GTP hydrolysis. By combining NMR, molecular dynamics simulations, kinetic assays, and mutational analysis, we find that the PH domain binds Arf·GTP at the membrane, to establish an active state primed for GTP hydrolysis. We identify key residues on the PH domain and Arf that drive this allosteric mechanism, which mathematical modeling shows contributes as much to GTPase activation as membrane recruitment. The finding that PH domains directly modulate small GTPases has broad implications for the Ras and Rho oncoprotein families.

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Metadata

Authors
Soubias, O., Foley, S. L., Jian, X., Jackson, R. A., Zhang, Y., Rosenberg, E. M., Hu, B. J., Li, J., Heinrich, F., Johnson, M. E., Sodt, A. J., Randazzo, P. A., & Byrd, R. A.
Affiliations
  • National Institutes of Health (US)
  • Johns Hopkins University (US)
  • National Cancer Institute (US)
  • National Institute of Standards and Technology (US)
Corresponding author
  • Randazzo, Paul A.
  • Byrd, R. Andrew
ORCID
Venue
Nature Communications
Publisher
Springer Science and Business Media LLC
Published
2025-09-30
Volume
16
Issue
1
Pages
8701-8701
ISSN
2041-1723 (ISSN-L)
DOI
10.1038/s41467-025-63764-w
PMC
PMC12485133
OpenAlex
W4414650702
Cited by
1
References
1
Citations by year
2026: 1
FWCI
0.30
Type
journal article
Language
en
Open access
Licence
Locations
  • Nature Communications (publishedVersion, cc-by)
  • Research Square (acceptedVersion, cc-by)
  • Europe PMC (PubMed Central) (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • PubMed Central (publishedVersion, cc-by)
  • Europe PMC (PubMed Central) (submittedVersion, cc-by)
  • PubMed Central (submittedVersion, cc-by)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
Funders
  • U.S. Department of Health & Human Services | NIH | National Cancer Institute (ZIABC007365, ZIA BC011131)
  • U.S. Department of Health & Human Services | NIH | National Institute of General Medical Sciences (R35GM133644)
  • U.S. Department of Health & Human Services | NIH | Eunice Kennedy Shriver National Institute of Child Health and Human Development (ZIA HD008955)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Kinase Regulation and GTPase Signaling
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Cellular transport and secretion
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Receptor Mechanisms and Signaling
Keywords
ASAP1, Arf1, GTPase-activating protein, GTP hydrolysis, pleckstrin homology domain, allosteric mechanism, membrane recruitment, small GTPase regulation, oncogenic Ras, tumor progression

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