Raf-1 Kinase Possesses Distinct Binding Domains for Phosphatidylserine and Phosphatidic Acid

Ghosh, Sujoy; Strum, Jay C.; Sciorra, Vicki A.; Daniel, Larry W.; Bell, Robert M.

doi:10.1074/jbc.271.14.8472

Cited by 374 publications

(244 citation statements)

References 62 publications

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“…Inspection of the NMR structure of the RafCRD (Mott et al, 1996) suggests that a likely binding site for PS includes Raf-1 residues 128 ± 147, which form a hydrophobic channel. This is consistent with the data of Ghosh et al, 1994;1996 who showed that two Raf-1 fragments, each containing the region 128 ± 147 of the RafCRD bound PS equally well. It is also worth noting that residues 143 and 144 probably comprise part of the 14-3-3 binding site (Clark et al, 1997) and that overlapping binding sites for PS/Ca and 14-3-3 would o er a simple competitive mechanism for the displacement of 14-3-3 by PS/Ca.…”

Section: Discussionsupporting

confidence: 92%

“…Since both PS and 14-3-3 bind to the RafCRD (Ghosh et al, 1994;Clark et al, 1997), we investigated to what extent the binding of 14-3-3 to Raf-1 was in¯uenced by PS. A mixture of calcium, diacylglycerol and PS has been shown previously to activate Raf-1 in vitro (Force et al, 1994).…”

Section: Resultsmentioning

confidence: 99%

“…These data indicate that an intact zinc ®nger is required for some aspect of Raf-1 activation that occurs at the plasma membrane. In addition to Ras, two other CRD ligands have been identi®ed: 14-3-3 proteins, which bind to two phosphoserine-containing sites on Raf-1 (Muslin et al, 1996) as well as to the RafCRD (Michaud et al, 1995;Clark et al, 1997), and phosphatidylserine (PS), a plasma membrane inner lea¯et phospholipid (Ghosh et al, 1994) that can activate Raf-1 in vitro (Force et al, 1994).…”

Section: Introductionmentioning

confidence: 99%

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Interactions of c-Raf-1 with phosphatidylserine and 14-3-3

et al. 1999

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Section: Discussionsupporting

confidence: 92%

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Interactions of c-Raf-1 with phosphatidylserine and 14-3-3

et al. 1999

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“…GST fusion protein system is a useful system for analysis of ion binding to proteins and their domains. GST fusion protein approach was also utilized to identify and characterize Zn 2* binding to Raf-1 kinase [27] and to LIM homodomain protein Isl-2 [28].…”

Section: Discussionmentioning

confidence: 99%

Identification of the Zn²⁺ binding region in calreticulin

et al. 1995

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Calreticulin binds Zn 2+ with the relatively high affinit)/low capacity. To determine the location of the Zn 2+ binding site in calreticulin different domains of the protein were expressed in E. coli, using the glutathione S-transferase fusion protein system, and their Zn2+-dependent interaction with Zn2+-lDA-agarose were determined. Three distinct domains were used in this study: the N + P-domain (the first 290 residues); the N-domain (residues 1-182) and the proline-rich P-domain (residues 180-273). The N + P-domain bound to the Zn2+-lDA-agarose and were eluted with an increasing concentration of imidazole. The N-domain also bound 65Zn2+ as measured by the overlay method. The P-domain did not interact with the ZnZ+-IDA-agarose and it did not bind any detectable amount of Zu 2+. Chemical modification of calreticulin with diethyl pyrocarbonate indicated that rise out of seven histidines were protected in the presence of Zn 2+ but they were modified by diethyl pyrocarbonate in the absence of Zn 2+ suggesting that these residues may be involved in Zn 2+ binding to ealreticulin. We conclude that Zn 2+ binding sites in calreticulin are localized to the N-domain of the protein, region that is not involved in Ca 2+ binding to calreticulin.

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“…Yet another level of complexity was added to the regulation of Raf-1 when it was demonstrated that the cysteine-rich domain (CRD) of Raf-1 binds to the anionic membrane phospholipid, phosphatidylserine (PS) (Ghosh et al, 1994). Moreover, it has now been demonstrated that the addition of PS to a cellular preparation of K-Ras and B-Raf enhances the activation of B-Raf and that the addition of a mixture of plasma membrane lipid comonents, including PS, allows Ras to activate c-Raf-1 in vitro (Stokoe and McCormick, 1997).…”

Section: Introductionmentioning

confidence: 99%

Increasing complexity of Ras signaling

et al. 1998

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The initial discovery that ras genes endowed retroviruses with potent carcinogenic properties and the subsequent determination that mutated ras genes were present in a wide variety of human cancers, prompted a strong suspicion that the growth-promoting actions of mutated Ras proteins contribute to their aberrant regulation of growth stimulatory signaling pathways. In 1993, a remarkable convergence of experimental observations from genetic analyses of Drosophila, S. cerevisiae and C. elegans as well as biochemical and biological studies in mammalian cells came together to de®ne a clear role for Ras in signal transduction. What emerged was an elegant linear signaling pathway where Ras functions as a relay switch that is positioned downstream of cell surface receptor tyrosine kinases and upstream of a cytoplasmic cascade of kinases that included the mitogen-activated protein kinases (MAPKs). Activated MAPKs in turn regulated the activities of nuclear transcription factors. Thus, a signaling cascade where every component between the cell surface and the nucleus was de®ned and conserved in worms,¯ies and man. This was a remarkable achievement in our e orts to appreciate how the aberrant function of Ras proteins may contribute to the malignant growth properties of the cancer cell. However, the identi®cation of this pathway has proven to be just the beginning, rather than the culmination, of our understanding of Ras in signal transduction. Instead, we now appreciate that this simple linear pathway represents but a minor component of a very complex signaling circuitry. Ras signaling has emerged to involve a complex array of signaling pathways, where cross-talk, feedback loops, branch points and multi-component signaling complexes are recurring themes. The simplest concept of a signaling cascade, where each component simply relays the same message to the next, is clearly not the case. In this review, we summarize our current understanding of Ras signal transduction with an emphasis on new complexities associated with the recognition and/or activation of cellular e ectors, and the diverse array of signaling pathways mediated by interaction between Ras and Ras-subfamily proteins with multiple e ectors.

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Raf-1 Kinase Possesses Distinct Binding Domains for Phosphatidylserine and Phosphatidic Acid

Cited by 374 publications

References 62 publications

Interactions of c-Raf-1 with phosphatidylserine and 14-3-3

Interactions of c-Raf-1 with phosphatidylserine and 14-3-3

Identification of the Zn²⁺ binding region in calreticulin

Increasing complexity of Ras signaling

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Raf-1 Kinase Possesses Distinct Binding Domains for Phosphatidylserine and Phosphatidic Acid

Cited by 374 publications

References 62 publications

Interactions of c-Raf-1 with phosphatidylserine and 14-3-3

Interactions of c-Raf-1 with phosphatidylserine and 14-3-3

Identification of the Zn2+ binding region in calreticulin

Increasing complexity of Ras signaling

Contact Info

Product

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Identification of the Zn²⁺ binding region in calreticulin