title: Exchange protein directly activated by cAMP 1 (Epac1) is expressed in human neutrophils and mediates cAMP-dependent activation of the monomeric GTPase Rap1.
abstract: Epac1 and Epac2 bind cAMP and mediate cAMP-dependent activation of Rap1. cAMP is produced in neutrophils in response to many chemoattractants. This second messenger plays a key role in the regulation of the functions of neutrophils. However, it is still not known whether Epacs are expressed in human neutrophils. We found that stimulation of PLB-985 cells differentiated into neutrophil-like cells, human neutrophils with 8CPT-2Me-cAMP (a selective activator of Epacs), or FK (a diterpene that augments the intracellular level of cAMP) led to GTP-loading of Rap1. Epac1 mRNA was expressed in UND and DF PLB-985 cells, but Epac1 protein was only detected in DF PLB-985 cells. In human neutrophils, the Epac1 transcript was present, and Epac1 protein could be detected by Western blot analysis if the cells had been treated with the serine protease inhibitor PMSF. FK induced adhesion of PLB-985 cells and human neutrophils on fibrinogen, a ligand for β2 integrins. Interestingly, in DF PLB-985 cells, but not in human neutrophils, 8CPT-2Me-cAMP induced β2 integrin-dependent adhesion. The failure of 8CPT-2Me-cAMP to induce β2 integrin-dependent human neutrophil adhesion could be explained by the fact that this compound did not induce a switch of the β2 integrins from a low-affinity to a high-affinity ligand-binding conformation. We concluded that Epac1 is expressed in human neutrophils and is involved in cAMP-dependent regulation of Rap1. However, the loading of GTP on Rap1 per se is not sufficient to promote activation of β2 integrins.
authors.affiliation: Centre for Infection and Immunity, Queen’s University of Belfast, Belfast, Northern Ireland, United Kingdom.
authors.forename: Madhuri
authors.initials: M
authors.lastname: Dash-Koney
authors.forename: Ravi K
authors.initials: RK
authors.lastname: Deevi
authors.forename: Cheryl
authors.initials: C
authors.lastname: McFarlane
authors.forename: Karim
authors.initials: K
authors.lastname: Dib
mesh_terms: D018821:CD18 Antigens / Q000378:metabolism; D002448:Cell Adhesion / Q000187:drug effects / Q000502:physiology; D002460:Cell Line; D000242:Cyclic AMP / Q000378:metabolism*; D005786:Gene Expression Regulation / Q000187:drug effects / Q000502:physiology*; D020662:Guanine Nucleotide Exchange Factors / Q000096:biosynthesis*; D006801:Humans; D009504:Neutrophils / Q000166:cytology / Q000378:metabolism*; D015842:Serine Proteinase Inhibitors / Q000494:pharmacology; D020827:rap1 GTP-Binding Proteins / Q000378:metabolism*
chemical_list: D018821:CD18 Antigens; D020662:Guanine Nucleotide Exchange Factors; C116122:RAPGEF3 protein, human; D015842:Serine Proteinase Inhibitors; D000242:Cyclic AMP; D020827:rap1 GTP-Binding Proteins