Rap guanine nucleotide exchange factor (GEF) 4 (RAPGEF4), also known as exchange protein directly activated by cAMP 2 (EPAC2) is a
protein that in humans is encoded by the RAPGEF4gene.[5][6][7]
Epac2 is a target of
cAMP, a major
second messenger in various cells. Epac2 is coded by the RAPGEF4 gene, and is expressed mainly in brain, neuroendocrine, and endocrine tissues.[8] Epac2 functions as a guanine nucleotide exchange factor for the Ras-like small GTPase Rap upon cAMP stimulation.[8][9] Epac2 is involved in a variety of cAMP-mediated cellular functions in endocrine and neuroendocrine cells and neurons.[10][11]
Gene and transcripts
Human Epac2 is coded by RAPGEF4 located at chromosome 2q31-q32, and three
isoforms (Epac2A, Epac2B, and Epac2C) are generated by alternate promoter usage and differential splicing.[8][12][13] Epac2A (called Epac2 originally) is a multi-domain protein with 1,011 amino acids, and is expressed mainly in brain and
neuroendocrine and
endocrine tissues such as
pancreatic islets and neuroendocrine cells.[8] Epac2A is composed of two regions, an
amino-terminal regulatory region and a
carboxy-terminal catalytic region. The regulatory region contains two
cyclic nucleotide-binding domains (cNBD-A and cNBD-B) and a
DEP (
Dishevelled,
Egl-10, and
Pleckstrin) domain. The catalytic region, which is responsible for the activation of Rap, consists of a CDC25 homology domain (CDC25-HD), a Ras exchange motif (REM) domain, and a Ras association (RA) domain.[14] Epac2B is devoid of the first cNBD-A domain and Epac2C is devoid of a cNBD-A and a DEP domain. Epac2B and Epac2C are expressed specifically in adrenal gland[12] and liver,[13] respectively.
Mechanism of action
The crystal structure reveals that the catalytic region of Epac2 interacts with cNBD-B intramolecularly, and in the absence of cAMP is sterically masked by a regulatory region, which thereby inhibits interaction between the catalytic region and
Rap1.[15] The crystal structure of the cAMP analog-bound active form of Epac2 in a complex with Rap1B indicates that the binding of cAMP to the cNBD-B domain induces the dynamic conformational changes that allow the regulatory region to rotate away. This conformational change enables access of Rap1 to the catalytic region and allows activation.[15][16]
Specific agonists
Several Epac-selective cAMP analogs have been developed to clarify the functional roles of Epacs as well those of the Epac-dependent signaling pathway distinct from the
PKA-dependent signaling pathway.[17] The modifications of 8-position in the purine structure and 2’-position in ribose is considered to be crucial to the specificity for Epacs. So far, 8-pCPT-2’-O-Me-cAMP (8-pCPT) and its membrane permeable form 8-pCPT-AM are used for their great specificity toward Epacs.
Sulfonylurea drugs (SUs), widely used for the treatment of type 2 diabetes through stimulation of insulin secretion from pancreatic β-cells, have also been shown to specifically activate Epac2.[18]
Function
In
pancreatic β-cells, cAMP signaling, which can be activated by various extracellular stimuli including hormonal and neural inputs primarily through Gs-coupled receptors, is of importance for normal regulation of insulin secretion to maintain glucose homeostasis. Activation of cAMP signaling amplifies insulin secretion by Epac2-dependent as well as PKA-dependent pathways.[10] Epac2-Rap1 signaling is critical to promote exocytosis of insulin-containing vesicles from the readily releasable pool.[19] In Epac2-mediated
exocytosis of insulin granules, Epac2 interacts with Rim2,[20][21] which is a scaffold protein localized in both plasma membrane and insulin granules, and determines the docking and priming states of exocytosis.[22][23] In addition,
piccolo, a possible Ca2+ sensor protein,[24] interacts with the Epac2-Rim2 complex to regulate cAMP-induced insulin secretion.[22] It is suggested that
phospholipase C-ε (PLC-ε), one of the effector proteins of Rap, regulates intracellular Ca2+ dynamics by altering the activities of ion channels such as ATP-sensitive potassium channel, ryanodine receptor, and IP3 receptor.[11][25]
In neurons, Epac is involved in
neurotransmitter release in
glutamatergicsynapses from
calyx of Held and in crayfish neuromuscular junction.[26][27][28] Epac also has roles in the development of brain by regulation of neurite growth and neuronal differentiation as well as axon regeneration in mammalian tissue.[29][30] Furthermore, Epac2 may regulate synaptic plasticity, and thus control higher brain functions such as memory and learning.[31][32]
In heart, Epac1 is expressed predominantly, and is involved in the development of hypertrophic events by chronic cAMP stimulation through
β-adrenergic receptors.[33] In contrast, chronic stimulation of Epac2 may be a cause of
cardiac arrhythmia through
CaMKII-dependent
diastolicsarcoplasmic reticulum (SR) Ca2+ release in mice.[34][35] Epac2 also is involved in
GLP-1-stimulated
atrial natriuretic peptide (ANP) secretion from heart.[36]
Clinical implications
As Epac2 is involved in many physiological functions in various cells, defects in the Epac2/Rap1 signaling mechanism could contribute to the development of various pathological states. Studies of Epac2
knockout mice indicate that Epac-mediated signaling is required for potentiation of insulin secretion by
incretins (gut hormones released from enteroendocrine cells following meal ingestion) such as
glucagon-like peptide-1 (GLP-1) and
glucose-dependent insulinotropic polypeptide,[37][38] suggesting that Epac2 is a promising target for treatment of diabetes. In fact, incretin-based diabetes therapies are currently used in clinical practice worldwide; development of Epac2-selective agonists might well lead to the discovery of further novel anti-diabetic drugs. An analog of GLP-1 has been shown to exert a blood pressure-lowering effect by stimulation of atrial natriuretic peptide (ANP) secretion through Epac2.[36] In heart, chronic stimulation of β-adrenergic receptor is known to progress to arrhythmia through an Epac2-dependent mechanism.[34][35] In brain, up-regulation of Epac1 and down-regulation of Epac2 mRNA are observed in patients with
Alzheimer's disease, suggesting roles of Epacs in the disease.[39] An Epac2 rare coding variant is found in patients with autism and could be responsible for the dendritic morphological abnormalities.[40][41]
Thus, Epac2 is involved in the pathogenesis and pathophysiology of various diseases, and represents a promising therapeutic target.
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