Erratum: Errata

Cheever, Matthew L.; Sato, Trey K.; Beer, Tonny de; Kutateladze, Tatiana G.; Emr, Scott D.; Overduin, Michael

doi:10.1038/35083007

Cited by 39 publications

(70 citation statements)

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“…Protein-lipid overlay assays were performed as described (20). In brief, membrane arrays (PIPStrips) were purchased from Echelon Research Laboratories (Salt Lake City, UT).…”

Section: Expression and Purification Of Glutathione S-transferase (Gst)-mentioning

confidence: 99%

PIP ₃ inhibition of RGS protein and its reversal by Ca ²⁺ /calmodulin mediate voltage-dependent control of the G protein cycle in a cardiac K ⁺ channel

Ishii

Inanobe

Kurachi

2002

Proc. Natl. Acad. Sci. U.S.A.

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Regulators of G protein signaling (RGS) accelerate intrinsic GTP hydrolysis on ␣ subunits of trimeric G proteins and play crucial roles in the physiological regulation of G protein-mediated cell signaling. The control mechanisms of the action of RGS proteins per se are poorly clarified, however. We recently showed a physiological mode of action of a RGS protein in cardiac myocytes. The voltagedependent formation of Ca 2؉ ͞calmodulin facilitated the GTPase activity of RGS by an unidentified mechanism, which underlay the ''relaxation'' behavior of G protein-gated K ؉ (KG) channels. Here we report the mechanism which is the reversal by Ca 2؉ ͞calmodulin of phosphatidylinositol-3,4,5,-trisphosphate (PIP3)-mediated inhibition of RGS. Purified RGS4 protein alone inhibited GTP-induced KG channel activity in inside-out patches from atrial myocytes. The inhibitory effect of RGS4 was reduced by PIP3 and restored by addition of Ca 2؉ ͞calmodulin. The intracellular application of anti-PIP3 antibody abolished the RGS-dependent relaxation behavior of KG current in atrial myocytes. This study, therefore, reveals a general physiological control mechanism of RGS proteins by lipidprotein interaction. H eterotrimeric G proteins mediate different intracellular signaling cascades and regulate many cellular functions (1). Although numerous kinds of G protein-coupled receptors and effectors have been identified, the existence of regulators of the G protein cycle has been neglected for a long period. Recently, a family of cytosolic proteins named ''regulators of G protein signaling'' (RGS proteins) has been identified (2, 3). These proteins share the conserved ''RGS domain'' of Ϸ120 aa which is responsible for accelerating GTPase activity on the G protein ␣ subunit (4-6). RGS proteins are thought to play a central role in the physiological regulation of the G protein cycle, and their importance has been confirmed in the immune response (7) and sensory perception (8, 9). To date, more than 20 mammalian RGS proteins have been identified. These RGS proteins vary in their molecular structure, tissue distribution, and intracellular localization, and thus may play divergent functional roles in different tissues.We recently found that RGS proteins were responsible not only for the acceleration of the deactivation time course upon agonist washout (10), but also for the characteristic gating behavior of G protein-activated inward-rectifier K ϩ channels (K G ) in cardiac atrial myocytes (11-13). K G channels are directly activated by the ␤␥ subunits released from pertussis toxinsensitive G proteins upon receptor stimulation, and contribute to acetylcholine (ACh)-induced deceleration of heart beat and neurotransmitter-evoked slow inhibitory postsynaptic potentials in neurons (14). When membrane potential is suddenly hyperpolarized from positive potentials, the K G current in myocytes first instantaneously and then slowly increases. The latter timedependent current change is termed ''relaxation'' and is characteristic for native K G current (15). ...

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“…Protein-lipid overlay assays were performed as described (20). In brief, membrane arrays (PIPStrips) were purchased from Echelon Research Laboratories (Salt Lake City, UT).…”

Section: Expression and Purification Of Glutathione S-transferase (Gst)-mentioning

confidence: 99%

PIP ₃ inhibition of RGS protein and its reversal by Ca ²⁺ /calmodulin mediate voltage-dependent control of the G protein cycle in a cardiac K ⁺ channel

Ishii

Inanobe

Kurachi

2002

Proc. Natl. Acad. Sci. U.S.A.

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“…Although the majority of these domains can interact with several PIs, the FYVE domain is remarkably selective for PtdIns(3)P (7)(8)(9). In addition to PI ligation, these domains often insert hydrophobic elements into the membrane bilayer, as has been demonstrated for the C2 (10), ENTH (11), FERM (12), FYVE (13), and PX (14) domains and the vinculin tail (15). Insertion into the membrane can be accompanied by interactions with multiple lipid head groups.…”

mentioning

confidence: 99%

Multivalent Mechanism of Membrane Insertion by the FYVE Domain

Kutateladze

Capelluto

Ferguson

et al. 2004

Journal of Biological Chemistry

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109

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Targeting of a wide variety of proteins to membranes involves specific recognition of phospholipid head groups and insertion into lipid bilayers. For example, proteins that contain FYVE domains are recruited to endosomes through interaction with phosphatidylinositol 3-phosphate (PtdIns(3)P). However, the structural mechanism of membrane docking and insertion by this domain remains unclear. Here, the depth and angle of micelle insertion and the lipid binding properties of the FYVE domain of early endosome antigen 1 are estimated by NMR spectroscopy. Spin label probes incorporated into micelles identify a hydrophobic protuberance that inserts into the micelle core and is surrounded by interfacially active polar residues. A novel proxyl PtdIns(3)P derivative is developed to map the position of the phosphoinositide acyl chains, which are found to align with the membrane insertion element. Dual engagement of the FYVE domain with PtdIns(3)P and dodecylphosphocholine micelles yields a 6-fold enhancement of affinity. The additional interaction of phosphatidylserine with a conserved basic site of the protein further amplifies the micelle binding affinity and dramatically alters the angle of insertion. Thus, the FYVE domain is targeted to endosomes through the synergistic action of stereospecific PtdIns(3)P head group ligation, hydrophobic insertion and electrostatic interactions with acidic phospholipids.Cellular processes including signal transduction, vesicular trafficking, and cytoskeletal rearrangement require selective recruitment of proteins to membrane surfaces. Well established mechanisms for localizing cytosolic proteins to membranes include electrostatic interactions through a basic peptide sequence, anchoring by covalently attached acyl chains, and association with the cytoplasmic domains of transmembrane proteins (reviewed in Refs. 1 and 2). (4), PDZ (5), and PTB (6) domains. Although the majority of these domains can interact with several PIs, the FYVE domain is remarkably selective for PtdIns(3)P (7-9). In addition to PI ligation, these domains often insert hydrophobic elements into the membrane bilayer, as has been demonstrated for the C2 (10), ENTH (11), FERM (12), FYVE (13), and PX (14) domains and the vinculin tail (15). Insertion into the membrane can be accompanied by interactions with multiple lipid head groups. For example, the PX domain of the p47 subunit of the NADPH oxidase binds cooperatively to PtdIns(3)P and phosphatidic acid (16), and the vinculin tail co-ligates phosphatidylinositol 4,5-bisphosphate and PtdSer (15). Although it is becoming evident that insertion of proteins into membranes is widespread, the three-dimensional orientations and quantitative binding properties remain challenging to characterize. The most common electron paramagnetic resonance and fluorescence approaches have provided important insights (17-20) but require covalent attachment of paramagnetic groups to various positions of the protein or mutations of residues. The inevitable effects of these modifications on lipi...

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“…These effectors are characterized by the presence of specialized phosphoinositide-binding domains. These domains include the pleckstrin homology (PH) domain (6), the FYVE finger (7), and the recently discovered PHOX homology (PX) domain (8). Furthermore, C2 domains have been shown to bind to phospholipids or phosphoinositides in the presence or absence of calcium (9).…”

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confidence: 99%

The C2A domain of JFC1 binds to 3′-phosphorylated phosphoinositides and directs plasma membrane association in living cells

Catz

Johnson

Babior

2002

Proc. Natl. Acad. Sci. U.S.A.

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Phosphatidylinositol 3-kinase products play a central role in the regulation of several intracellular pathways via adaptor proteins that share the ability to bind to 3 -phosphoinositides with high affinity and specificity. JFC1 is a C2 domain-containing protein involved in cellular trafficking that has been shown to bind 3 -phosphoinositides in vitro. In this work, we demonstrate that the C2A domain of JFC1 is the module responsible for its binding to the plasma membrane via 3 -phosphoinositides in vivo. We show that the C2A domain of JFC1 is the only domain present in this protein that localizes to the plasma membrane in living cells. Moreover, the C2A domain of JFC1 binds 3 -phosphoinositides in vitro with similar specificity as that described for full-length JFC1, suggesting that the domain mediates the specific membrane localization of the full-length protein. Furthermore, the C2A domain of JFC1 colocalized with the pleckstrin homology domain of Akt in vivo, and both the JFC1 C2A domain and the full-length JFC1 dissociated from the membrane in the presence of PI 3-kinase specific inhibitors. We also show that the association of the C2A domain to the membrane is modulated by calcium. From these results we analyze possible mechanisms for the role of JFC1 in cellular trafficking. P hosphoinositides are specialized phospholipids known to localize to the inner leaflet of the plasma membrane, vesicles, and the nuclear envelope and participate in several signaling pathways (1). One of these phosphoinositides, phosphatidylinositol (4,5)-bisphosphate [PtdIns(4,5)P 2 ], plays a central role in the regulation of several intracellular pathways, from actin cytoskeleton modulation and vesicle trafficking (2) to modulation of the transcription factor Tubby protein (3). Furthermore, PtdIns(4,5)P 2 is a precursor for the second messenger phosphatidylinositol (3,4,5) trisphosphate [PtdIns(3,4,5)P 3 ] via the phosphoinositide 3-kinases ref. 4). PI 3-kinases and their products 3Ј-phosphorylated phosphoinositides (3Ј-phosphoinositides) are also involved in many crucial cell functions including vesicle trafficking (1, 2), cytoskeleton modulation, and cell survival (5). It is now clear that most of the functions regulated by phosphoinositides are mediated by adaptor proteins that share the ability to bind to PtdIns(4,5)P 2 and PI 3-kinase products with high affinity and specificity. These effectors are characterized by the presence of specialized phosphoinositide-binding domains. These domains include the pleckstrin homology (PH) domain (6), the FYVE finger (7), and the recently discovered PHOX homology (PX) domain (8). Furthermore, C2 domains have been shown to bind to phospholipids or phosphoinositides in the presence or absence of calcium (9). Proteins that contain C2 domains could generally be categorized as signal transduction mediators or membrane traffic effectors (10). The most extensively studied proteins included in the last group are the synaptotagmins (Syts). Syts are a group of transmembrane proteins that contain tan...

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Erratum: Errata

Abstract: Vesicle-mediated protein transport: regulatory interactions between the Vps15 protein kinase and the Vps34 PtdIns 3-kinase essential for protein sorting to the vacuole in yeast.

Cited by 39 publications

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PIP ₃ inhibition of RGS protein and its reversal by Ca ²⁺ /calmodulin mediate voltage-dependent control of the G protein cycle in a cardiac K ⁺ channel

PIP ₃ inhibition of RGS protein and its reversal by Ca ²⁺ /calmodulin mediate voltage-dependent control of the G protein cycle in a cardiac K ⁺ channel

Multivalent Mechanism of Membrane Insertion by the FYVE Domain

The C2A domain of JFC1 binds to 3′-phosphorylated phosphoinositides and directs plasma membrane association in living cells

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Erratum: Errata

Abstract: Vesicle-mediated protein transport: regulatory interactions between the Vps15 protein kinase and the Vps34 PtdIns 3-kinase essential for protein sorting to the vacuole in yeast.

Cited by 39 publications

References 0 publications

PIP 3 inhibition of RGS protein and its reversal by Ca 2+ /calmodulin mediate voltage-dependent control of the G protein cycle in a cardiac K + channel

PIP 3 inhibition of RGS protein and its reversal by Ca 2+ /calmodulin mediate voltage-dependent control of the G protein cycle in a cardiac K + channel

Multivalent Mechanism of Membrane Insertion by the FYVE Domain

The C2A domain of JFC1 binds to 3′-phosphorylated phosphoinositides and directs plasma membrane association in living cells

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Product

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PIP ₃ inhibition of RGS protein and its reversal by Ca ²⁺ /calmodulin mediate voltage-dependent control of the G protein cycle in a cardiac K ⁺ channel

PIP ₃ inhibition of RGS protein and its reversal by Ca ²⁺ /calmodulin mediate voltage-dependent control of the G protein cycle in a cardiac K ⁺ channel