title: Trafficking of GFP-AQP5 chimeric proteins conferred with unphosphorylated amino acids at their PKA-target motif ((152)SRRTS) in MDCK-II cells.
abstract: Three constructs having mutated PKA-target motif at (152)SRRTS of AQP5, an exocrine type water channel, were prepared and fused to C-terminus of green fluorescence protein cDNA to examine the effects of blocking of phosphorylation at (152)SRRTS (a consensus PKA-target motif of AQP5) on translocation or trafficking of the chimeric proteins expressed in the Madin-Darby canine kidney-II (MDCK-II) cells. H-89 treatment increased translocation of wild-type GFP-AQP5 to the apical membrane. All 3 mutant molecules translocated 1.5 to 2 times more than the control wild-type GFP-AQP5. Colchicine but not cytochalasin B inhibited the translocation of wild-type GFP-AQP5. Present results suggest dephosphorylation of this consensus sequence increase GFP-AQP5 translocation, and that microtubules but not microfilaments are involved in this event.
authors.affiliation: Department of Molecular Oral Physiology, Institute of Health Biosciences, the University of Tokushima Graduate School, Japan.
authors.forename: Mileva Ratko
authors.initials: MR
authors.lastname: Karabasil
authors.forename: Takahiro
authors.initials: T
authors.lastname: Hasegawa
authors.forename: Ahmad
authors.initials: A
authors.lastname: Azlina
authors.forename: Nunuk
authors.initials: N
authors.lastname: Purwanti
authors.forename: Javkhlan
authors.initials: J
authors.lastname: Purevjav
authors.forename: Chenjuan
authors.initials: C
authors.lastname: Yao
authors.forename: Tetsuya
authors.initials: T
authors.lastname: Akamatsu
authors.forename: Kazuo
authors.initials: K
authors.lastname: Hosoi
mesh_terms: D020816:Amino Acid Motifs; D000596:Amino Acids / Q000378:metabolism*; D000818:Animals; D051402:Aquaporin 5 / Q000235:genetics / Q000378:metabolism*; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D002678:Chimera / Q000235:genetics; D003078:Colchicine / Q000494:pharmacology; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism*; D003571:Cytochalasin B / Q000494:pharmacology; D004285:Dogs; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism*; D007546:Isoquinolines / Q000494:pharmacology; D007668:Kidney / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D010766:Phosphorylation; D021381:Protein Transport; D013449:Sulfonamides / Q000494:pharmacology
chemical_list: D000596:Amino Acids; D051402:Aquaporin 5; D007546:Isoquinolines; D013449:Sulfonamides; D049452:Green Fluorescent Proteins; D003571:Cytochalasin B; D017868:Cyclic AMP-Dependent Protein Kinases; C063509:N-(2-(4-bromocinnamylamino)ethyl)-5-isoquinolinesulfonamide; D003078:Colchicine