Crystal structure of the pleckstrin homology domain from dynamin

Timm, David E.; Salim, Kamran; Gout, Ivan; Guruprasad, Lalitha; Waterfield, Mike; Blundell, Tom L.

doi:10.1038/nsb1194-782

Cited by 120 publications

(81 citation statements)

References 28 publications

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“…Interestingly, a variety of PH domains from various other signaling molecules that were bacterially expressed and purified under similar conditions did not show any biological effect when injected in the oocytes, suggesting that the observed effect is specific to the Sos1 PH domain. Different PH domains appear to share a common tertiary structure, although the conservation of their primary sequence is rather limited (2)(3)(4)(5)(6). In this regard, the present results suggest that, in addition to the tertiary structure, the primary structure is an essential factor responsible for the observed biological activity.…”

Section: Sos1 Ph Cooperates With Normal Ras Proteins and Is Blocked Bmentioning

confidence: 57%

“…Included among the many PH-containing proteins described so far are several that participate in Ras signaling pathways. Despite having a rather limited primary sequence consensus identity, the PH domains from different proteins appear to share a common tertiary structure consisting of seven antiparallel ␤-sheets and a carboxyl-terminal amphiphilic ␣ helix (3)(4)(5)(6).…”

mentioning

confidence: 99%

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Isolated Sos1 PH Domain Exhibits Germinal Vesicle Breakdown-inducing Activity in Oocytes

Mora

Guerrero

Mahadevan

et al. 1996

Journal of Biological Chemistry

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Purified, bacterially expressed PH domains of Sos1, IRS-1, ␤ARK, and PLC␦ 1 were analyzed functionally by means of microinjection into full grown, stage VI Xenopus laevis oocytes. Whereas the PH domains from IRS-1, ␤ARK, or PLC␦ 1 did not show any effect in the oocytes, injection of the purified Sos1 PH domain resulted in induction of significant rates of germinal vesicle breakdown and meiotic maturation. Furthermore, the Sos1 PH domain exhibited also significant synergy with insulin or coinjected normal Ras protein in induction of germinal vesicle breakdown, although it did not affect the rate of progesterone-induced maturation. These results suggest that purified, isolated PH domains retain, at least in part, their functional specificity and that Xenopus oocytes may constitute a useful biological system to analyze the functional role of the Sos1 PH domain in Ras signaling pathways. Pleckstrin homology (PH)1 domains are modular domains of about 100 amino acids present in a variety of signaling and cytoskeletal proteins, which were initially defined by their homology with the internal repeats of pleckstrin, a major protein kinase C substrate in platelets (1, 2). Included among the many PH-containing proteins described so far are several that participate in Ras signaling pathways. Despite having a rather limited primary sequence consensus identity, the PH domains from different proteins appear to share a common tertiary structure consisting of seven antiparallel ␤-sheets and a carboxyl-terminal amphiphilic ␣ helix (3-6).The functional role of PH domains remains unclear. Some reports describe a role of PH domains in protein-protein interactions such as Akt homooligomerization (7) and association with G␤␥ subunits (8 -10) or protein kinase C (11, 12), whereas others point to interactions with membrane lipids such as phosphatidylinositol 4,5-bisphosphate (13-17). An interesting hypothesis suggests that all those different interactions may represent alternative methods for directing the implicated proteins to membrane locations. Genetic evidence has confirmed the functional importance of PH domains since there is an established association between specific point mutations in the PH domain of Bruton tyrosine kinase and X-chromosome linked human agammaglobulinemia or mouse inmunodeficiency (18 -21). Regarding Ras signaling, recent reports point to the presence of positive regulatory elements in the region of the Ras guanine nucleotide exchanger Sos1 protein that encompasses its PH (and also the contiguous DH) domain (22).Xenopus oocytes provide a useful experimental model for the functional analysis of signaling molecules, which are able to induce their meiotic maturation, i.e. germinal vesicle breakdown (GVBD). Xenopus oocytes possess at least two independent pathways leading to GVBD. In one, progesterone leads to decreased adenylate cyclase activity, with a resulting drop in overall cAMP levels and protein kinase A-dependent phosphorylations. In the other, insulin or IGF-1 triggers a cascade of phosphorylations initi...

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Section: Sos1 Ph Cooperates With Normal Ras Proteins and Is Blocked Bmentioning

confidence: 57%

mentioning

confidence: 99%

Isolated Sos1 PH Domain Exhibits Germinal Vesicle Breakdown-inducing Activity in Oocytes

Mora

Guerrero

Mahadevan

et al. 1996

Journal of Biological Chemistry

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“…Although isolated Dyn1PH crystallized as a dimer (33,53), x-ray scattering (33), and analytical ultracentrifugation (54) studies showed that it is monomeric at concentrations up to 10 mg/ml (680 M), and tends to oligomerize only above 1 mM (54). Oligomerization of Dyn1PH therefore requires that it is present in a molecule that self-associates through other interactions.…”

Section: Discussionmentioning

confidence: 99%

The Pleckstrin Homology Domains of Dynamin Isoforms Require Oligomerization for High Affinity Phosphoinositide Binding

Klein

Lee

Frank³

et al. 1998

Journal of Biological Chemistry

193

181

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The dynamins are 100-kDa GTPases involved in the scission event required for formation of endocytotic vesicles. The two main described mammalian dynamins (dynamin؊1 and dynamin؊2) both contain a pleckstrin homology (PH) domain, which has been implicated in dynamin binding to (and activation by) acidic phospholipids, most notably phosphoinositides. We demonstrate that the PH domains of both dynamin isoforms require oligomerization for high affinity phosphoinositide binding. Strong phosphoinositide binding was detected only when the PH domains were dimerized by fusion to glutathione S-transferase, or via a single engineered intermolecular disulfide bond. Phosphoinositide binding specificities agreed reasonably with reported effects of different phospholipids on dynamin GTPase activity. Although they differ in their ability to inhibit rapid endocytosis in adrenal chromaffin cells, the dynamin؊1 and dynamin؊2 PH domains showed identical phosphoinositide binding specificities. Since oligomerization is required for binding of the dynamin PH domain to phosphoinositides, it follows that PH domain-mediated phosphoinositide binding will favor oligomerization of intact dynamin (which has an inherent tendency to self-associate). We propose that the dynamin PH domain thus mediates the observed cooperative binding of dynamin to membranes containing acidic phospholipids and promotes the self-assembly that is critical for both stimulation of its GTPase activity and its ability to achieve membrane scission.

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“…The structure of several PH domains has been determined (Ferguson et al, 1994(Ferguson et al, , 1995Fushman et al, 1995;Hyvonen et al, 1995;Macias et al, 1994;Timm et al, 1994;Yoon et al, 1994). These structures exhibit a common globular fold that consists of seven anti-parallel b strands and an a helix (Gibson et al, 1994;Lemmon et al, 1996).…”

Section: A Structural Model Of the Akt Ph Domainmentioning

confidence: 99%

Akt activation by growth factors is a multiple-step process: the role of the PH domain

et al. 1998

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The protein kinase encoded by the Akt proto-oncogene is activated by phospholipid binding, membrane translocation and phosphorylation. To address the relative roles of these mechanisms of Akt activation, we have employed a combination of genetic and pharmacological approaches. Transient transfection of NIH3T3 cells with wild-type Akt, pleckstrin homology (PH) domain mutants, generated on the basis of a PH domain structural model, and phosphorylation site Akt mutants provided evidence for a model of Akt activation consisting of three sequential steps: (1) a PH domain-dependent, growth factor-independent step, marked by constitutive phosphorylation of threonine 450 (T450) and perhaps serine 124 (S124), that renders the protein responsive to subsequent activation events; (2) a growth factor-induced, PI3-K-dependent membrane-translocation step; and (3) a PI3-K-dependent step, characterized by phosphorylation at T308 and S473, that occurs in the cell membrane and is required for activation. When forced to translocate to the membrane, wild-type Akt and PH domain Akt mutants that are defective in the ®rst step become constitutively active, suggesting that the purpose of this step is to prepare the protein for membrane translocation. Both growth factor stimulation and forced membrane translocation, however, failed to activate a T308A mutant. This, combined with the ®nding that T308D/S473D double mutant is constitutively active, suggests that the purpose of the three-step process of Akt activation is the phosphorylation of the protein at T308 and S473. The proposed model provides a framework for a comprehensive understanding of the temporal and spatial requirements for Akt activation by growth factors.

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Crystal structure of the pleckstrin homology domain from dynamin

Cited by 120 publications

References 28 publications

Isolated Sos1 PH Domain Exhibits Germinal Vesicle Breakdown-inducing Activity in Oocytes

Isolated Sos1 PH Domain Exhibits Germinal Vesicle Breakdown-inducing Activity in Oocytes

The Pleckstrin Homology Domains of Dynamin Isoforms Require Oligomerization for High Affinity Phosphoinositide Binding

Akt activation by growth factors is a multiple-step process: the role of the PH domain

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