CoChat
Sign Up
Cited by 13Receptor Mechanisms and SignalingProtein Kinase Regulation and GTPase SignalingNeuroscience and Neuropharmacology Research

The DOI matches its Crossref record; 2 of the 2 references it cites were found in a registry or an index.

Dissecting the molecular basis for the modulation of neurotransmitter GPCR signaling by GINIP

Open access
Luebbers, Alex, Gonzalez-Hernandez, Alberto J., Zhou, Myles, Eyles, Stephen J., Levitz, Joshua, Garcia-Marcos, Mikel·2024·Structure
DOI 10.1016/j.str.2023.10.010PMC PMC10872408

Published version of a preprint

Open this paper in CoChat — free account

Abstract

It is well established that G-protein-coupled receptors (GPCRs) stimulated by neurotransmitters are critical for neuromodulation. Much less is known about how heterotrimeric G-protein (Gαβγ) regulation after receptor-mediated activation contributes to neuromodulation. Recent evidence indicates that the neuronal protein GINIP shapes GPCR inhibitory neuromodulation via a unique mechanism of G-protein regulation that controls pain and seizure susceptibility. However, the molecular basis of this mechanism remains ill-defined because the structural determinants of GINIP responsible for binding and regulating G proteins are not known. Here, we combined hydrogen-deuterium exchange mass spectrometry, computational structure predictions, biochemistry, and cell-based biophysical assays to demonstrate an effector-like binding mode of GINIP to Gαi. Specific amino acids of GINIP's PHD domain first loop are essential for G-protein binding and subsequent regulation of Gαi-GTP and Gβγ signaling upon neurotransmitter GPCR stimulation. In summary, these findings shed light onto the molecular basis for a post-receptor mechanism of G-protein regulation that fine-tunes inhibitory neuromodulation.

Overview

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Integrity signals

Registry

Crossref · 10.1016/j.str.2023.10.010

Cites

2 of 2 references verified0 citation errors · 0 not checkable

Statistics

Verification

Claims the paper makes

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

The self-check needs the paper’s full text.

Citations

Cites

Research works printed but not linked: 73

  1. Roth, 2019, Molecular pharmacology of metabotropic receptors targeted by neuropsychiatric drugs, Nat. Struct. Mol. Biol., 26, 535, 10.1038/s41594-019-0252-8 · DOI
  2. Greengard, 2001, The neurobiology of slow synaptic transmission, Science, 294, 1024, 10.1126/science.294.5544.1024 · DOI
  3. Zurawski, 2019, The expanding roles and mechanisms of G protein-mediated presynaptic inhibition, J. Biol. Chem., 294, 1661, 10.1074/jbc.TM118.004163 · DOI
  4. Hauser, 2017, Trends in GPCR drug discovery: new agents, targets and indications, Nat. Rev. Drug Discov., 16, 829, 10.1038/nrd.2017.178 · DOI
  5. Hopkins, 2002, The druggable genome, Nat. Rev. Drug Discov., 1, 727, 10.1038/nrd892 · DOI · Paper page
  6. Santos, 2017, A comprehensive map of molecular drug targets, Nat. Rev. Drug Discov., 16, 19, 10.1038/nrd.2016.230 · DOI · Paper page
  7. Sriram, 2018, G Protein-Coupled Receptors as Targets for Approved Drugs: How Many Targets and How Many Drugs?, Mol. Pharmacol., 93, 251, 10.1124/mol.117.111062 · DOI · Paper page
  8. Gilman, 1987, G proteins: transducers of receptor-generated signals, Annu. Rev. Biochem., 56, 615, 10.1146/annurev.bi.56.070187.003151 · DOI
  9. Ulrich, 2007, GABA(B) receptors: synaptic functions and mechanisms of diversity, Curr. Opin. Neurobiol., 17, 298, 10.1016/j.conb.2007.04.001 · DOI
  10. Gurevich, 2016, G protein-coupled receptor kinases as regulators of dopamine receptor functions, Pharmacol. Res., 111, 1, 10.1016/j.phrs.2016.05.010 · DOI
  11. Bylund, 1994, International Union of Pharmacology nomenclature of adrenoceptors, Pharmacol. Rev., 46, 121, 10.1016/S0031-6997(25)06780-8 · DOI
  12. Betke, 2012, GPCR mediated regulation of synaptic transmission, Prog. Neurobiol., 96, 304, 10.1016/j.pneurobio.2012.01.009 · DOI
  13. Csanády, 2017, A new target for G protein signaling, Elife, 6, 10.7554/eLife.31106 · DOI
  14. Smrcka, 2019, G-protein betagamma subunits as multi-functional scaffolds and transducers in G-protein-coupled receptor signaling, Cell. Mol. Life Sci., 76, 4447, 10.1007/s00018-019-03275-2 · DOI
  15. Berman, 1996, GAIP and RGS4 are GTPase-activating proteins for the Gi subfamily of G protein alpha subunits, Cell, 86, 445, 10.1016/S0092-8674(00)80117-8 · DOI
  16. Watson, 1996, RGS family members: GTPase-activating proteins for heterotrimeric G-protein alpha-subunits, Nature, 383, 172, 10.1038/383172a0 · DOI
  17. Dohlman, 1997, RGS proteins and signaling by heterotrimeric G proteins, J. Biol. Chem., 272, 3871, 10.1074/jbc.272.7.3871 · DOI
  18. De Vries, 1996, GAIP is membrane-anchored by palmitoylation and interacts with the activated (GTP-bound) form of G alpha i subunits, Proc. Natl. Acad. Sci. USA, 93, 15203, 10.1073/pnas.93.26.15203 · DOI
  19. Soundararajan, 2008, Structural diversity in the RGS domain and its interaction with heterotrimeric G protein alpha-subunits, Proc. Natl. Acad. Sci. USA, 105, 6457, 10.1073/pnas.0801508105 · DOI
  20. Hepler, 1999, Emerging roles for RGS proteins in cell signalling, Trends Pharmacol. Sci., 20, 376, 10.1016/S0165-6147(99)01369-3 · DOI
  21. Han, 2006, RGS2 determines short-term synaptic plasticity in hippocampal neurons by regulating Gi/o-mediated inhibition of presynaptic Ca2+ channels, Neuron, 51, 575, 10.1016/j.neuron.2006.07.012 · DOI
  22. Anderson, 2010, R7BP complexes with RGS9-2 and RGS7 in the striatum differentially control motor learning and locomotor responses to cocaine, Neuropsychopharmacology, 35, 1040, 10.1038/npp.2009.212 · DOI
  23. Zhou, 2012, GIRK channel modulation by assembly with allosterically regulated RGS proteins, Proc. Natl. Acad. Sci. USA, 109, 19977, 10.1073/pnas.1214337109 · DOI
  24. Neubig, 2002, Regulators of G-Protein signalling as new central nervous system drug targets, Nat. Rev. Drug Discov., 1, 187, 10.1038/nrd747 · DOI
  25. Gerber, 2016, Roles for Regulator of G Protein Signaling Proteins in Synaptic Signaling and Plasticity, Mol. Pharmacol., 89, 273, 10.1124/mol.115.102210 · DOI
  26. Park, 2023, Fine-tuning GPCR-mediated neuromodulation by biasing signaling through different G protein subunits, Mol. Cell, 83, 2540, 10.1016/j.molcel.2023.06.006 · DOI
  27. Gaillard, 2014, GINIP, a Galphai-interacting protein, functions as a key modulator of peripheral GABAB receptor-mediated analgesia, Neuron, 84, 123, 10.1016/j.neuron.2014.08.056 · DOI
  28. Englander, 2006, Hydrogen exchange and mass spectrometry: A historical perspective, J. Am. Soc. Mass Spectrom., 17, 1481, 10.1016/j.jasms.2006.06.006 · DOI
  29. Engen, 2015, Analytical Aspects of Hydrogen Exchange Mass Spectrometry, Annu. Rev. Anal. Chem., 8, 127, 10.1146/annurev-anchem-062011-143113 · DOI
  30. Hvidt, 1966, Hydrogen exchange in proteins, Adv. Protein Chem., 21, 287, 10.1016/S0065-3233(08)60129-1 · DOI
  31. James, 2022, Advances in Hydrogen/Deuterium Exchange Mass Spectrometry and the Pursuit of Challenging Biological Systems, Chem. Rev., 122, 7562, 10.1021/acs.chemrev.1c00279 · DOI
  32. Masson, 2019, Recommendations for performing, interpreting and reporting hydrogen deuterium exchange mass spectrometry (HDX-MS) experiments, Nat. Methods, 16, 595, 10.1038/s41592-019-0459-y · DOI
  33. Suzuki, 2010, Strategy for comprehensive identification of human N-myristoylated proteins using an insect cell-free protein synthesis system, Proteomics, 10, 1780, 10.1002/pmic.200900783 · DOI
  34. Mirdita, 2022, ColabFold: making protein folding accessible to all, Nat. Methods, 19, 679, 10.1038/s41592-022-01488-1 · DOI · Paper page
  35. Sprang, 1997, G protein mechanisms: Insights from Structural Analysis, Annu. Rev. Biochem., 66, 639, 10.1146/annurev.biochem.66.1.639 · DOI
  36. Johnston, 2006, Minimal determinants for binding activated G alpha from the structure of a G alpha(i1)-peptide dimer, Biochemistry, 45, 11390, 10.1021/bi0613832 · DOI
  37. Dai, 2022, State-selective modulation of heterotrimeric Galphas signaling with macrocyclic peptides, Cell, 185, 3950, 10.1016/j.cell.2022.09.019 · DOI
  38. Won, 2023, Molecular architecture of the Gα(i)-bound TRPC5 ion channel, Nat. Commun., 14, 2550, 10.1038/s41467-023-38281-3 · DOI
  39. Qi, 2022, Structural basis of adenylyl cyclase 9 activation, Nat. Commun., 13, 1045, 10.1038/s41467-022-28685-y · DOI
  40. Slep, 2001, Structural determinants for regulation of phosphodiesterase by a G protein at 2.0 A, Nature, 409, 1071, 10.1038/35059138 · DOI
  41. Lyon, 2013, Full-length Galpha(q)-phospholipase C-beta3 structure reveals interfaces of the C-terminal coiled-coil domain, Nat. Struct. Mol. Biol., 20, 355, 10.1038/nsmb.2497 · DOI
  42. Waldo, 2010, Kinetic scaffolding mediated by a phospholipase C-beta and Gq signaling complex, Science (New York, N.Y, 330, 974, 10.1126/science.1193438 · DOI
  43. Dessauer, 1998, Identification of a Gialpha binding site on type V adenylyl cyclase, J. Biol. Chem., 273, 25831, 10.1074/jbc.273.40.25831 · DOI
  44. Grishina, 1997, Identification of common and distinct residues involved in the interaction of alphai2 and alphas with adenylyl cyclase, J. Biol. Chem., 272, 20619, 10.1074/jbc.272.33.20619 · DOI
  45. Tesmer, 1997, Structure of RGS4 bound to AlF4--activated G(i alpha1): stabilization of the transition state for GTP hydrolysis, Cell, 89, 251, 10.1016/S0092-8674(00)80204-4 · DOI
  46. Kuramoto, 2017, Identification of Candidate Genes for Generalized Tonic-Clonic Seizures in Noda Epileptic Rat, Behav. Genet., 47, 609, 10.1007/s10519-017-9870-2 · DOI
  47. Li, 2006, Molecular basis for site-specific read-out of histone H3K4me3 by the BPTF PHD finger of NURF, Nature, 442, 91, 10.1038/nature04802 · DOI
  48. Peña, 2006, Molecular mechanism of histone H3K4me3 recognition by plant homeodomain of ING2, Nature, 442, 100, 10.1038/nature04814 · DOI
  49. Fiedler, 2008, Decoding of methylated histone H3 tail by the Pygo-BCL9 Wnt signaling complex, Mol. Cell, 30, 507, 10.1016/j.molcel.2008.03.011 · DOI
  50. Miller, 2010, Allosteric remodelling of the histone H3 binding pocket in the Pygo2 PHD finger triggered by its binding to the B9L/BCL9 co-factor, J. Mol. Biol., 401, 969, 10.1016/j.jmb.2010.07.007 · DOI
  51. Sanchez, 2011, The PHD finger: a versatile epigenome reader, Trends Biochem. Sci., 36, 364
  52. Musselman, 2011, Handpicking epigenetic marks with PHD fingers, Nucleic Acids Res., 39, 9061, 10.1093/nar/gkr613 · DOI
  53. Amato, 2018, Targeting Ligandable Pockets on Plant Homeodomain (PHD) Zinc Finger Domains by a Fragment-Based Approach, ACS Chem. Biol., 13, 915, 10.1021/acschembio.7b01093 · DOI
  54. Nakata, 2005, Functional characterization of Galphao signaling through G protein-regulated inducer of neurite outgrowth 1, Mol. Pharmacol., 67, 695, 10.1124/mol.104.003913 · DOI
  55. Mototani, 2018, Role of G protein-regulated inducer of neurite outgrowth 3 (GRIN3) in β-arrestin 2-Akt signaling and dopaminergic behaviors, Pflugers Arch., 470, 937, 10.1007/s00424-018-2124-1 · DOI
  56. Iida, 2004, Identification and biochemical analysis of GRIN1 and GRIN2, Methods Enzymol., 390, 475, 10.1016/S0076-6879(04)90029-8 · DOI
  57. Serikawa, 2019, Increased seizure sensitivity, emotional defects and cognitive impairment in PHD finger protein 24 (Phf24)-null rats, Behav. Brain Res., 369, 10.1016/j.bbr.2019.111922 · DOI
  58. Stols, 2002, A new vector for high-throughput, ligation-independent cloning encoding a tobacco etch virus protease cleavage site, Protein Expr. Purif., 25, 8, 10.1006/prep.2001.1603 · DOI
  59. Garcia-Marcos, 2009, GIV is a nonreceptor GEF for G alpha i with a unique motif that regulates Akt signaling, Proc. Natl. Acad. Sci. USA, 106, 3178, 10.1073/pnas.0900294106 · DOI
  60. Garcia-Marcos, 2020, Optogenetic activation of heterotrimeric G-proteins by LOV2GIVe, a rationally engineered modular protein, Elife, 9, 10.7554/eLife.60155 · DOI
  61. Marivin, 2020, DAPLE protein inhibits nucleotide exchange on Gα(s) and Gα(q) via the same motif that activates Gαi, J. Biol. Chem., 295, 2270, 10.1074/jbc.RA119.011648 · DOI
  62. Mumby, 1994, Myristoylation of G-protein alpha subunits, Methods Enzymol., 237, 254, 10.1016/S0076-6879(94)37067-2 · DOI
  63. Garcia-Marcos, 2010, A structural determinant that renders G alpha(i) sensitive to activation by GIV/girdin is required to promote cell migration, J. Biol. Chem., 285, 12765, 10.1074/jbc.M109.045161 · DOI
  64. Masuho, 2015, Distinct profiles of functional discrimination among G proteins determine the actions of G protein-coupled receptors, Sci. Signal., 8, ra123, 10.1126/scisignal.aab4068 · DOI
  65. Hollins, 2009, The c-terminus of GRK3 indicates rapid dissociation of G protein heterotrimers, Cell. Signal., 21, 1015, 10.1016/j.cellsig.2009.02.017 · DOI
  66. Vivaudou, 1997, Probing the G-protein regulation of GIRK1 and GIRK4, the two subunits of the KACh channel, using functional homomeric mutants, J. Biol. Chem., 272, 31553, 10.1074/jbc.272.50.31553 · DOI
  67. Leyme, 2017, Specific inhibition of GPCR-independent G protein signaling by a rationally engineered protein, Proc. Natl. Acad. Sci. USA, 114, E10319, 10.1073/pnas.1707992114 · DOI
  68. Pettersen, 2004, UCSF Chimera--a visualization system for exploratory research and analysis, J. Comput. Chem., 25, 1605, 10.1002/jcc.20084 · DOI
  69. Maziarz, 2018, A biochemical and genetic discovery pipeline identifies PLCdelta4b as a nonreceptor activator of heterotrimeric G-proteins, J. Biol. Chem., 293, 16964, 10.1074/jbc.RA118.003580 · DOI
  70. Garcia-Marcos, 2021, Complementary biosensors reveal different G-protein signaling modes triggered by GPCRs and non-receptor activators, Elife, 10, 10.7554/eLife.65620 · DOI
  71. DiGiacomo, 2020, Probing the mutational landscape of regulators of G protein signaling proteins in cancer, Sci. Signal., 13, 10.1126/scisignal.aax8620 · DOI
  72. Park, 2023, Fine-tuning GPCR-mediated neuromodulation by biasing signaling through different G-protein subunits, bioRxiv
  73. Boyer, 2009, Direct interaction of GABAB receptors with M2 muscarinic receptors enhances muscarinic signaling, J. Neurosci., 29, 15796, 10.1523/JNEUROSCI.4103-09.2009 · DOI

Metadata

Authors
Luebbers, A., Gonzalez-Hernandez, A. J., Zhou, M., Eyles, S. J., Levitz, J., & Garcia-Marcos, M.
Affiliations
  • Boston University (US)
  • Weill Cornell Medicine (US)
  • University of Massachusetts Amherst (US)
ORCID
Venue
Structure
Publisher
Elsevier BV
Published
2024-01
Volume
32
Issue
1
Pages
47-59.e7
ISSN
0969-2126 (ISSN-L)
DOI
10.1016/j.str.2023.10.010
PMC
PMC10872408
OpenAlex
W4388824607
Cited by
13
References
2
Citations by year
2026: 3 · 2025: 1 · 2024: 8
FWCI
1.16
Type
journal article
Language
en
Open access
Licence
Locations
  • Structure (publishedVersion)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by-nc)
  • PubMed (publishedVersion)
  • PubMed Central (acceptedVersion)
  • PubMed Central (submittedVersion, cc-by-nc)
Funders
  • Texas A&M University Health Science Center
  • National Institutes of Health (F31NS115318, R01NS117101)
  • Israel National Road Safety Authority (SCR_019063)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Receptor Mechanisms and Signaling
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Kinase Regulation and GTPase Signaling
  • Life Sciences › Neuroscience › Cellular and Molecular Neuroscience › Neuroscience and Neuropharmacology Research
Keywords
G protein-coupled receptors, G protein regulation, Gαi, Gβγ signaling, neurotransmitter signaling, PHD domain, G protein binding, pain susceptibility, seizure susceptibility, post-receptor signaling

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

Are you an author, or do you see a problem with this page? Request a review