In vitro processing of recombinant G protein gamma subunits. Requirements for assembly of an active beta gamma complex.

Higgins, Joyce B.; Casey, Patrick J.

doi:10.1016/s0021-9258(17)37077-1

Cited by 109 publications

(14 citation statements)

References 34 publications

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“…Although in platelets two different groups of proteins with known signal transduction properties have been shown to undergo prenylation and methylation (small GTP-binding proteins and γ subunits of heterotrimeric G-proteins), we suggest that, as in other cells [12][13][14][15], the effects observed on SRCE were due entirely to the inactivation of small GTP-binding proteins by FC analogues. Previous studies have suggested that prenylation might be important for the interaction between βγ complexes and the α subunit but that it is not required for βγ assembly [50,51]. However, it has recently been demonstrated that the prenylation of γ subunits of heterotrimeric G-proteins has only a negligible effect on signal transduction processes, which can be correlated with a decrease in hydrophobicity of the βγ subunits [52].…”

Section: Discussionmentioning

confidence: 99%

Farnesylcysteine analogues inhibit store-regulated Ca2+ entry in human platelets: evidence for involvement of small GTP-binding proteins and actin cytoskeleton

Rosado¹,

Sage²

2000

Biochem. J.

106

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We have investigated the mechanism of Ca(2+) entry into fura-2-loaded human platelets by preventing the prenylation of proteins such as small GTP-binding proteins. The farnesylcysteine analogues farnesylthioacetic acid (FTA) and N-acetyl-S-geranylgeranyl-L-cysteine (AGGC), which are inhibitors of the methylation of prenylated and geranylgeranylated proteins respectively, significantly decreased thrombin-evoked increases in intracellular free Ca(2+) concentration ([Ca(2+)](i)) in the presence, but not in the absence, of external Ca(2+), suggesting a relatively selective inhibition of Ca(2+) entry over internal release. Both these compounds and N-acetyl-S-farnesyl-L-cysteine, which had similar effects to those of FTA, also decreased Ca(2+) entry evoked by the depletion of intracellular Ca(2+) stores with thapsigargin. The inactive control N-acetyl-S-geranyl-L-cysteine was without effect. Patulin, an inhibitor of prenylation that is inert with respect to methyltransferases, also decreased store-regulated Ca(2+) entry. Cytochalasin D, an inhibitor of actin polymerization, significantly decreased store-regulated Ca(2+) entry in a time-dependent manner. Both cytochalasin D and the farnesylcysteine analogues FTA and AGGC inhibited actin polymerization; however, when evoking the same extent of decrease in actin filament formation, FTA and AGGC showed greater inhibitory effects on Ca(2+) entry, indicating a cytoskeleton-independent component in the regulation of Ca(2+) entry by small GTP-binding-protein. These findings suggest that prenylated proteins such as small GTP-binding proteins are involved in store-regulated Ca(2+) entry through actin cytoskeleton-dependent and cytoskeleton-independent mechanisms in human platelets.

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Section: Discussionmentioning

confidence: 99%

Farnesylcysteine analogues inhibit store-regulated Ca2+ entry in human platelets: evidence for involvement of small GTP-binding proteins and actin cytoskeleton

Rosado¹,

Sage²

2000

Biochem. J.

106

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“…29 Therefore, it is likely that the interaction between Gb 4 and Ga, but not the interaction between Gb 4 and Gg, is affected in both the p.Gly53Asp and p.Lys89Glu Gb 4 altered proteins. Although the dimerization of Gb and Gg is obligatory in nature, 31 the interaction between Gg and the altered forms of Gb 4 might render a decrease in the functional wild-type Gb 4 -Gg dimer and thus exert a dominant-negative effect that abolishes relevant GPCR signaling.…”

mentioning

confidence: 99%

Exome Sequencing Identifies GNB4 Mutations as a Cause of Dominant Intermediate Charcot-Marie-Tooth Disease

Soong

Huang²,

Tsai³

et al. 2013

The American Journal of Human Genetics

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Charcot-Marie-Tooth disease (CMT) is a heterogeneous group of inherited neuropathies. Mutations in approximately 45 genes have been identified as being associated with CMT. Nevertheless, the genetic etiologies of at least 30% of CMTs have yet to be elucidated. Using a genome-wide linkage study, we previously mapped a dominant intermediate CMT to chromosomal region 3q28-q29. Subsequent exome sequencing of two affected first cousins revealed heterozygous mutation c.158G>A (p.Gly53Asp) in GNB4, encoding guanine-nucleotide-binding protein subunit beta-4 (Gβ4), to cosegregate with the CMT phenotype in the family. Further analysis of GNB4 in an additional 88 unrelated CMT individuals uncovered another de novo mutation, c.265A>G (p.Lys89Glu), in this gene in one individual. Immunohistochemistry studies revealed that Gβ4 was abundant in the axons and Schwann cells of peripheral nerves and that expression of Gβ4 was significantly reduced in the sural nerve of the two individuals carrying the c.158G>A (p.Gly53Asp) mutation. In vitro studies demonstrated that both the p.Gly53Asp and p.Lys89Glu altered proteins impaired bradykinin-induced G-protein-coupled-receptor (GPCR) signaling, which was facilitated by the wild-type Gβ4. This study identifies GNB4 mutations as a cause of CMT and highlights the importance of Gβ4-related GPCR signaling in peripheral-nerve function in humans.

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“…Therefore, complexing Gβγ constitutes a second membrane docking signal for singly modified Gα subunits [21]. Prenylation of the Gγ subunit is required not only for proper Gβγ membrane localization but also for effective heterotrimer Gαβγ formation and for particular interaction with effector proteins such as adenylyl cyclase or phospholipase Cβ2 [86,87]. Farnesyl anchors drive membrane proteins to non-raft domains [88,89].…”

Section: Prenylationmentioning

confidence: 99%

Every Detail Matters. That Is, How the Interaction between Gα Proteins and Membrane Affects Their Function

2021

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In highly organized multicellular organisms such as humans, the functions of an individual cell are dependent on signal transduction through G protein-coupled receptors (GPCRs) and subsequently heterotrimeric G proteins. As most of the elements belonging to the signal transduction system are bound to lipid membranes, researchers are showing increasing interest in studying the accompanying protein–lipid interactions, which have been demonstrated to not only provide the environment but also regulate proper and efficient signal transduction. The mode of interaction between the cell membrane and G proteins is well known. Despite this, the recognition mechanisms at the molecular level and how the individual G protein-membrane attachment signals are interrelated in the process of the complex control of membrane targeting of G proteins remain unelucidated. This review focuses on the mechanisms by which mammalian Gα subunits of G proteins interact with lipids and the factors responsible for the specificity of membrane association. We summarize recent data on how these signaling proteins are precisely targeted to a specific site in the membrane region by introducing well-defined modifications as well as through the presence of polybasic regions within these proteins and interactions with other components of the heterocomplex.

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In vitro processing of recombinant G protein gamma subunits. Requirements for assembly of an active beta gamma complex.

Cited by 109 publications

References 34 publications

Farnesylcysteine analogues inhibit store-regulated Ca2+ entry in human platelets: evidence for involvement of small GTP-binding proteins and actin cytoskeleton

Farnesylcysteine analogues inhibit store-regulated Ca2+ entry in human platelets: evidence for involvement of small GTP-binding proteins and actin cytoskeleton

Exome Sequencing Identifies GNB4 Mutations as a Cause of Dominant Intermediate Charcot-Marie-Tooth Disease

Every Detail Matters. That Is, How the Interaction between Gα Proteins and Membrane Affects Their Function

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