Trafficking of GFP-AQP5 chimeric proteins conferred with unphosphorylated amino acids at their PKA-target motif ((152)SRRTS) in MDCK-II cells.
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.
Karabasil
Mileva Ratko
MR
Department of Molecular Oral Physiology, Institute of Health Biosciences, the University of Tokushima Graduate School, Japan.
Hasegawa
Takahiro
T
Azlina
Ahmad
A
Purwanti
Nunuk
N
Purevjav
Javkhlan
J
Yao
Chenjuan
C
Akamatsu
Tetsuya
T
Hosoi
Kazuo
K
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
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