Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts:   Target Concepts:
Query: EC:2.7.11.25 (MEKK1)
 1,856 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Eukaryotic cells respond to hyperosmotic conditions by expunging water from the cell, leading to cell shrinkage. This is counteracted by adaptive responses that restore cell volume and strengthen the cytoskeletal architecture. In the budding yeast Saccharomyces cerevisiae, this response is mediated primarily by the mitogen-activated protein kinase (MAPK) cascade CDC42-STE50-STE11-Pbs2-Hog1. In mammalian cells, MAPK scaffold proteins facilitate the efficiency of signaling within the cascade by placing a kinase near its substrate and also regulate the subcellular localization of the signaling. Our laboratory has discovered a scaffold that coordinates the analogous Hog1 signal in mammalian cells, termed OSM (osmosensing scaffold for MEKK3). OSM organizes a complex consisting of the small GTPase Rac, MEKK3, and MKK3 for the activation of p38 MAPK. Interactions among OSM, Rac, and MEKK3 are augmented in response to sorbitol and are also localized to membrane ruffles, sites of rapid actin turnover. Suppression of the expression of OSM or MEKK3 by RNA interference strongly inhibits the sorbitol-dependent activation of p38. Furthermore, mutations in OSM were concurrently found to cause cerebral cavernous malformations (CCM), a disease of the central nervous system characterized by thin-walled, leaky blood vessels that become hemorrhagic. Our laboratory has also demonstrated that Krit1, another gene harboring mutations that lead to CCM, binds OSM and its interaction is enhanced in response to sorbitol in a similar manner as the MEKK3-OSM interaction. This chapter describes the cell biological and biochemical methods used for assaying protein-protein interactions in live cells using fluorescence resonance energy transfer, in vitro kinase assays for MEKK3-MKK3-p38 pathway members, and gene suppression by RNA interference to study hyperosmotic stress-dependent signaling.
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PMID:Hyperosmotic induction of mitogen-activated protein kinase scaffolding. 1787 25

Cerebral cavernous malformations (CCM) are sporadic or inherited vascular lesions of the central nervous system characterized by dilated, thin-walled, leaky vessels. Linkage studies have mapped autosomal dominant mutations to three loci: ccm1 (KRIT1), ccm2 (OSM), and ccm3 (PDCD10). All three proteins appear to be scaffolds or adaptor proteins, as no enzymatic function can be attributed to them. Our previous results demonstrated that OSM is a scaffold for the assembly of the GTPase Rac and the MAPK kinase kinase MEKK3, for the hyperosmotic stress-dependent activation of p38 MAPK. Herein, we show that the three CCM proteins are members of a larger signaling complex. To define this complex, epitope-tagged wild type OSM or OSM harboring the mutation of F217-->A, which renders the OSM phosphotyrosine binding (PTB) domain unable to bind KRIT1, were stably introduced into RAW264.7 mouse macrophages. FLAG-OSM or FLAG-OSMF217A and the associated complex members were purified by immunoprecipitation using anti-FLAG antibody. OSM binding partners were identified by gel-based methods combined with electrospray ionization-MS or by multidimensional protein identification technology (MudPIT). Previously identified proteins that associate with OSM including KRIT1, MEKK3, Rac, and the KRIT1-binding protein ICAP-1 were found in the immunoprecipitates. In addition, we show for the first time that PDCD10 binds to OSM and is found in cellular CCM complexes. Other prominent proteins that bound the CCM complex include EF1A1, RIN2, and tubulin, with each interaction disrupted with the OSMF217A mutant protein. We further show that PDCD10 binds phosphatidylinositol di- and triphosphates and OSM binds phosphatidylinositol monophosphates. The findings define the targeting of the CCM complex to membranes and to proteins regulating trafficking and the cytoskeleton.
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PMID:Proteomic identification of the cerebral cavernous malformation signaling complex. 1790 Jan 4

Cerebral cavernous malformations (CCM) are neurovascular dysplasias that result in mulberry-shaped lesions predominantly located in brain and spinal tissues. Mutations in three genes are associated with CCM. These genes encode for the proteins KRIT1/CCM1 (krev interaction trapped 1/cerebral cavernous malformations 1), cerebral cavernous malformations 2, osmosensing scaffold for MEKK3 (CCM2/malcavernin/OSM), and cerebral cavernous malformations 3/programmed cell death 10 (CCM3/PDCD10). There have been many significant recent advances in our understanding of the structure and function of these proteins, as well as in their roles in cellular signaling. Here, we provide an update on the current knowledge of the structure of the CCM proteins and their functions within cellular signaling, particularly in cellular adhesion complexes and signaling cascades. We go on to discuss subcellular localization of the CCM proteins, the formation and regulation of the CCM complex signaling platform, and current progress towards targeted therapy for CCM disease. Recent structural studies have begun to shed new light on CCM protein function, and we focus here on how these studies have helped inform the current understanding of these roles and how they may aid future studies into both CCM-related biology and disease mechanisms.
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PMID:Signaling pathways and the cerebral cavernous malformations proteins: lessons from structural biology. 2428 96

Cerebral cavernous malformations (CCM) are dysplasias that primarily occur in the neurovasculature, and are associated with mutations in three genes: KRIT1, CCM2, and PDCD10, the protein products of which are KRIT1 (Krev/Rap1 Interaction Trapped 1; CCM1, cerebral cavernous malformations 1), CCM2 (cerebral cavernous malformations 2; OSM, osmosensing scaffold for MEKK3), and CCM3 (cerebral cavernous malformations 3; PDCD10, programmed cell death 10). Until recently, these proteins were relatively understudied at the molecular level, and only three folded domains were documented. These were a band 4.1, ezrin, radixin, moesin (FERM), and an ankyrin repeat domain (ARD) in KRIT1, and a phosphotyrosine-binding (PTB) domain in CCM2. Over the past 10 years, a crystallographic approach has been used to discover a series of previously unidentified domains within the CCM proteins. These include a non-functional Nudix (or pseudonudix) domain in KRIT1, a harmonin homology domain (HHD) in CCM2, and dimerization and focal adhesion targeting (FAT)-homology domains within CCM3. Many of the roles of these domains have been revealed by structure-guided studies that show the CCM proteins can directly interact with one another to form a signaling scaffold, and that the "CCM complex" functions in signal transduction by interacting with other binding partners, including ICAP1, RAP1, and MEKK3. In this chapter, we describe the crystallization of CCM protein domains alone, and with their interaction partners.
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PMID:Crystallographic Studies of the Cerebral Cavernous Malformations Proteins. 3252 60