Expression of cGMP-dependent protein kinase in human atrium.
We have shown that cGMP-dependent protein kinase (PKG) mediates stimulation of L-type calcium current by cGMP in rabbit atrial myocytes. The human atrium may have similar PKG-dependent regulation of calcium current. To elucidate the significance of PKG in cardiac function, we have isolated human PKG type I alpha cDNA (+1 to 2016), determined the nucleotide sequence and analyzed specific expression of PKG in human atrium. We obtained full-length cDNA of PKG type I alpha from human atrial RNA using reverse transcriptase-polymerase chain reaction (RT-PCR). The coding region of human cardiac PKG I alpha showed 99.9% homology to previously published human PKG I alpha except for base No. 1983. At this position G was substituted for T and this resulted in an amino acid substitution from Leu649 to Phe649. The cloned PKG I alpha cDNA was expressed in COS cells and the expressed PKG showed cGMP-stimulated PKG enzyme activity and immunoreactivity. Ribonuclease protection assay, Western blot analysis, and PKG enzyme activity assays in homogenates from human atrial tissue demonstrated the presence of PKG mRNA and protein in human atrial tissue. Immunofluorescence staining confirmed that PKG is highly expressed in human atrial myocytes. These findings suggest that PKG is highly expressed in human atrium and that PKG-dependent phosphorylation may be important in regulation of calcium channel activity in human atrial myocytes.
Mishra
M
M
The Todd Franklin Cardiac Research Laboratory, The Sibley Children's Heart Center, Department of Pediatrics, 2040 Ridgewood Dr. NE, Atlanta, GA 30322, USA.
Wagner
M B
MB
Wang
Y
Y
Joyner
R W
RW
Kumar
R
R
D000328:Adult; D000368:Aged; D019943:Amino Acid Substitution; D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D019556:COS Cells; D017869:Cyclic GMP-Dependent Protein Kinases / Q000378:metabolism*; D005260:Female; D005455:Fluorescent Antibody Technique; D006325:Heart Atria / Q000201:enzymology*; D006801:Humans; D007527:Isoenzymes / Q000378:metabolism; D007700:Kinetics; D008297:Male; D018613:Microscopy, Confocal; D008875:Middle Aged; D011494:Protein Kinases / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012689:Sequence Homology, Nucleic Acid
D007527:Isoenzymes; D012333:RNA, Messenger; D011494:Protein Kinases; D017869:Cyclic GMP-Dependent Protein Kinases