Phosphoinositide 3-kinase family

PI3Ka Phosphoinositide 3-kinase family, accessory domain (PIK domain) E(MFP) 2(2) 3(3) 1QMM... 

Members of the phosphoinositide (PI)-3 kinase family appear to be involved in the phosphorylation of H2A.X. The SQ motif matches a common target site for these kinases and the formation of y-H2A.X in response to double stranded breaks is inhibited by wortmannin, an inhibitor of PI-3 kinases [63]. Examination of cell lines deficient in the PI-3 kinase ATM indicated that it has a major role in phosphorylating H2A.X in response to double strand breaks [64]. ATM can phosphorylate H2A.X in vitro suggesting that it may directly phosphorylate H2A.X in vivo [64]. Another PI-3 kinase ATR appears to be involved in phosphorylating H2A.X in response to replicational stress induced by treatment of dividing cells with hydroxyurea or by irradiating them with ultraviolet light [65]. It has been hypothesized that PI-3 kinases such as ATM are recruited to, or activated at, the site of the double stranded break and then phosphorylate H2A.X molecules around the break point [40,64,66]. 

In contrast to this polyspecific ABPP probe suitable for profiling a large fraction of the protein kinase family, several other approaches have been introduced, which focus on specific subfamilies. Wortmannin, for example, is a potent natural product specific for the inhibition of phosphoinositide 3- and polo-like kinases in vitro and in vivo. Its role in ABPP probe design will be discussed in the natural product-derived probe section (Section 9.17.3.3)97 98... 

Akt is activated by binding of plasma membrane phospholipids downstream of insulin receptors, growth and survival factor receptors in a phosphoinositide 3-kinases dependent manner. In humans, there are three genes in the Akt family Aktl, Akt2 and Akt3. Their respective fimctions are still under investigation. 

The family of heterotrimeric G proteins is involved in transmembrane signaling in the nervous system, with certain exceptions. The exceptions are instances of synaptic transmission mediated via receptors that contain intrinsic enzymatic activity, such as tyrosine kinase or guanylyl cyclase, or via receptors that form ion channels (see Ch. 10). Heterotrimeric G proteins were first identified, named and characterized by Alfred Gilman, Martin Rodbell and others close to 20 years ago. They consist of three distinct subunits, a, (3 and y. These proteins couple the activation of diverse types of plasmalemma receptor to a variety of intracellular processes. In fact, most types of neurotransmitter and peptide hormone receptor, as well as many cytokine and chemokine receptors, fall into a superfamily of structurally related molecules, termed G-protein-coupled receptors. These receptors are named for the role of G proteins in mediating the varied biological effects of the receptors (see Ch. 10). Consequently, numerous effector proteins are influenced by these heterotrimeric G proteins ion channels adenylyl cyclase phosphodiesterase (PDE) phosphoinositide-specific phospholipase C (PI-PLC), which catalyzes the hydrolysis of phosphatidylinositol 4,5-bisphosphate (PIP2) and phospholipase A2 (PLA2), which catalyzes the hydrolysis of membrane phospholipids to yield arachidonic acid. In addition, these G proteins have been implicated in... 

The inositol polyphosphate 5-phosphatases belong to a family of enzymes that terminate the signals generated by inositol lipid kinases and PLC. To date, two major types of 5-phosphatase have been identified, both of which share a common 5-phosphatase domain of approximately 300 amino acids, with several highly conserved motifs. Type-I enzymes are 43-65 kDa and preferentially hydrolyze 1(1,4,5)P3 and 1(1,3,4,5)P4, with the attendant formation of I(1,4)P2 and 1(1,3,4)P3, but have little or no activity towards membrane-bound phosphoinositides. The pro-totypic form of a type-15-phosphatase is a 43 kDa protein that is post-translationally modified by farnesylation of the carboxyl terminus CAAX motif this modification juxtaposes the enzyme with the membrane. Type-II enzymes are larger (75-160 kDa) and will hydrolyze both water-soluble inositol phosphates and lipids that... 

Phosphoinositide 3-kinases A conserved family of signal transducers. Trends Biochem. Sci. 22 267-272. 

MacDougall, E.K. Domin, J. Waterfield, M.D. A family of phosphoinositide 3-kinases in Drosophila identifies a new mediator of signal transduction. Curr. Biol., 5, 1404-1415 (1995)... 

Vanhaesebroeck, B. Leevers, S.J. Panayotou, G. Waterfield, M.D. Phosphoinositide 3-kinases a conserved family of signal transducers. Trends Biochem. Sci., 22, 267-272 (1997)... 

The probes of this type were shown to selectively label at least 75% of human kinases in crude cell lysates, thus demonstrating their selectivity and promiscuity for kinases [101]. As a follow up, the labeled kinases were subjected to proteolytic digestion, and the biotinylated peptides purified on avidin beads and analyzed by LC-MS/MS. This analysis demonstrated that the site of probe labeling was indeed the conserved active-site lysine as predicted. In contrast to the promiscuity demonstrated by the acyl phosphate probes, several selective covalent inhibitors of protein kinases have been used as ABPP probes. Wortmannin is a natural product derived from the fungus Penicillium funiculosum. It is a potent and specific covalent inhibitor of phosphoinositide 3-kinase (PI3K) and the PI3K-related kinase (PIKK) families [102, 103]. The use of natural products in relation to ABPP is covered by Breinbauer et al. [104]. 

Figure 4. Linear representation of the Arabidopsis PIPKs and PLCs. The Arabidopsis PtdlnsP 5-kinases are most similar to the human type I PtdlnsP 5-kinases. There are 11 putative type I /I/PtdlnsP 5-kinases in Arabidopsis arranged in two subfamilies based on size. Subfamily B contains X/PIPK1-9, all of which contain membrane occupation and recognition nexus (MORN) repeats. /1/PIPK10-11 are in Subfamily A with molecular weights less than that of the members of subfamily B and contain no MORN repeats. The Arabidopsis phosphoinositide specific phospholipase C family is most similar to the animal PLC . There are seven functional PI-PLCs in Arabidopisis (Hunt et al., 2004). All isoforms contain EF-hand motifs, the X and Y catalytic domains characteristic of PI-PLCs and a C2 lipid-binding domain.

PI-3 kinase Phosphoinositide 3-kinases a family of enzymes that are capable of phosphorylating the 3 position hydroxyl group of the inositol ring of phosphatidyiinositol. 

PLC)/Ca, and PKC signaling pathways in rat neutrophils [352]. Activation of phosphoinositide 3-kinase (PI3K) and Src family kinase is required for the respiratory burst in rat neutrophils stimulated with... 

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