Specificity and promiscuity in phosphoinositide binding by pleckstrin homology domains
Open accessAbstract
Pleckstrin homology (PH) domains are small protein modules involved in recruitment of signaling molecules to cellular membranes, in some cases by binding specific phosphoinositides. We describe use of a convenient "dot-blot" approach to screen 10 different PH domains for those that recognize particular phosphoinositides. Each PH domain bound phosphoinositides in the assay, but only two (from phospholipase C-delta1 and Grp1) showed clear specificity for a single species. Using soluble inositol phosphates, we show that the Grp1 PH domain (originally cloned on the basis of its phosphatidylinositol 3,4,5-trisphosphate (PtdIns(3,4,5)P3) binding) binds specifically to D-myo-inositol 1,3,4,5-tetrakisphosphate (Ins(1,3,4,5)P4) (the PtdIns(3,4,5)P3 headgroup) with KD = 27.3 nM, but binds D-myo-inositol 1,3,4-trisphosphate (Ins(1,3,4)P3) or D-myo-inositol 1,4,5-trisphosphate (Ins(1,4,5)P3) over 80-fold more weakly. We show that this specificity allows localization of the Grp1 PH domain to the plasma membrane of mammalian cells only when phosphatidylinositol 3-kinase (PI 3-K) is activated. The presence of three adjacent equatorial phosphate groups was critical for inositol phosphate binding by the Grp1 PH domain. By contrast, another PH domain capable of PI 3-K-dependent membrane recruitment (encoded by EST684797) does not distinguish Ins(1,3,4)P3 from Ins(1,3,4,5)P3 (binding both with very high affinity), despite selecting strongly against Ins(1,4,5)P3. The remaining PH domains tested appear significantly less specific for particular phosphoinositides. Together with data presented in the literature, our results suggest that many PH domains bind similarly to multiple phosphoinositides (and in some cases phosphatidylserine), and are likely to be regulated in vivo by the most abundant species to which they bind. Thus, using the same simple approach to study several PH domains simultaneously, our studies suggest that highly specific phosphoinositide binding is a characteristic of relatively few cases.
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The PH domain of PLCδ can also bind to phosphatidylinositol-3,4-bisphosphate and phosphatidylinositol-3,4,5-trisphosphate in vitro, though with comparatively low affinity.
“In vitro, the PH domain of PLCδ can also bind to phosphatidylinositol-3,4-bisphosphate (3,4-PIP2) and phosphatidylinositol-3,4,5-trisphosphate (3,4,5-PIP3), though with comparatively low affinity (Kavran et al. 1998).”
From Localized Biphasic Changes in Phosphatidylinositol-4,5-Bisphosphate at Sites of Phagocytosis (2000) · cites this paper as Kavran et al. 1998
The section on relative binding affinities says PLCδ-PH does not select against other phosphoinositides as strongly as some PH domains, which broadly supports binding beyond its preferred ligand. But the Discussion reports that, in vesicles containing 33% phosphatidylserine, PLCδ-PH bound PtdIns(3,4,5)P3 slightly more strongly than its preferred PtdIns(4,5)P2. The copy does not establish comparatively low affinity for both lipids named in the claim. (LLM, conf=0.82)
Experimental — accuracy not yet measuredThe GFP-tagged PH domains of Akt and Btk recognize specifically 3,4-PIP2 and 3,4,5-PIP3.
“To assess the relative abundance of these lipids, RAW macrophages were transfected with the GFP-tagged PH domains of either Akt (Fig. 1C and Fig. D) or Btk (not illustrated), which recognize specifically 3,4-PIP2 and 3,4,5-PIP3 (Kavran et al. 1998; Varnai et al. 1999).”
From Localized Biphasic Changes in Phosphatidylinositol-4,5-Bisphosphate at Sites of Phagocytosis (2000) · cites this paper as Kavran et al. 1998
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Research works printed but not linked: 5
- Yao, L., Kawakami, Y., and Kawakami, T. (1994) Proc. Natl. Acad. Sci. U. S. A. 91, 9175-9179
- Lemmon, M. A., Ferguson, K. M., O'Brien, R., Sigler, P. B., and Schlessinger, J. (1995) Proc. Natl. Acad. Sci. U. S. A. 92, 10472-10476
- Fushman, D., Cahill, S., Lemmon, M. A., Schlessinger, J., and Cowburn, D. (1995) Proc. Natl. Acad. Sci. U. S. A. 92, 816 -820
- Ausubel, F. M., Brent, R., Kingston, R. E., Moore, D. D., Seidman, J. G., Smith, J. A., Struhl, K., Albright, L. M., Coen, D. M., and Varki, A. (eds) (1997) Current Protocols in Molecular Biology, John Wiley & Sons, New York
- Margolis, B., and Young, R. A. (1995) in DNA Cloning 2 (Glover, D. M., and Hames, B. D., eds) pp. 1-14, IRL Press, Oxford
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- Authors
- Kavran, J. M., Klein, D. E., Lee, A., Falasca, M., Isakoff, S. J., Skolnik, E. Y., & Lemmon, M. A.
- Affiliations
- University of Pennsylvania (US)
- Mario Negri Institute for Pharmacological Research (IT)
- NYU Langone Health (US)
- Corresponding author
- Lemmon, Mark A.
- ORCID
- Venue
- Journal of Biological Chemistry
- Publisher
- Elsevier BV
- Published
- 1998-11
- Volume
- 273
- Issue
- 46
- Pages
- 30497-30508
- ISSN
- 0021-9258 (ISSN-L)
- OpenAlex
- W1972974751
- Cited by
- 434
- References
- 0
- Citations by year
- 2026: 8 · 2025: 8 · 2024: 1 · 2023: 6 · 2022: 8 · 2021: 7 · 2020: 7 · 2019: 11 · 2018: 10 · 2017: 5 · 2016: 6 · 2015: 14 · 2014: 17 · 2013: 11 · 2012: 15
- FWCI
- 9.44
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Licence
- Locations
- Journal of Biological Chemistry (publishedVersion, cc-by)
- PubMed (publishedVersion)
- Funders
- Damon Runyon Cancer Research Foundation
- University of Pennsylvania
- National Institutes of Health (2r01dk049207-05, DK49207)
- National Institute of Diabetes and Digestive and Kidney Diseases (DK 49207)
- 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 › PI3K/AKT/mTOR signaling in cancer
- Keywords
- pleckstrin homology domain, phosphoinositide binding, binding specificity, GRP1, phosphatidylinositol 3,4,5-trisphosphate, inositol tetrakisphosphate, phosphatidylinositol 3-kinase, membrane recruitment, phospholipase C delta 1, phosphatidylserine
- MeSH terms
- Animals, Binding Sites, Calorimetry, Humans, Inositol Phosphates, Isoenzymes, Kinetics, Ligands, Phosphatidylinositols, Type C Phospholipases, Structure-Activity Relationship, Sequence Homology, Amino Acid, Receptors, Cytoplasmic and Nuclear, Phosphatidylinositol 3-Kinases, Mice, Rats, Phospholipase C delta
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