A. Borges et al., Molecular and biochemical characterization of a CNP-Sensitive guanylyl cyclase in bovine tracheal smooth muscle, AM J RESP C, 25(1), 2001, pp. 98-103
Citations number
40
Categorie Soggetti
da verificare
Journal title
AMERICAN JOURNAL OF RESPIRATORY CELL AND MOLECULAR BIOLOGY
Muscarinic activation of bovine tracheal smooth muscle (BTSM) is involved i
n cyclic guanosine monophosphate (cGMP) production mediated through soluble
(sGC) and membrane-bound (mGC) guanylyl cyclases. A muscarinic- and NaCl-s
ensitive mGC exists in BTSM regulated by muscarinic receptors coupled to G
proteins. To identify the mGCs expressed in BTSM, reverse transcriptase/pol
ymerase chain reaction (RT-PCR) from total RNA was performed using degenera
te oligonucleotides for amplification of a region conserved among GC cataly
tic domains. Cloning of amplification products revealed that 76% of all BTS
M GC transcripts corresponded to the sGC beta1 subunit and 24% to the B-typ
e (C-type NP 1-22 [CNP]-sensitive) GC receptor. cGMP production by BTSM mem
brane and soluble fractions confirmed that sGC activity is 3-fold with resp
ect to mGC activity. RT-PCR using specific oligonucleotides revealed that A
(atrial NP-sensitive) and C (guanylin-sensitive) mGC subtypes are also exp
ressed in BTSM. Stimulation of basal plasma membrane GC activity by CNP was
higher than that by ANP, whereas guanylin showed no effect, indicating tha
t CNP-sensitive guanylyl cyclase (GC-B) is the predominant functional BTSM
mGC subtype. Strong adenosine triphosphate inhibition of CNP-stimulated mGC
activity supports the finding that the tracheal mGC isoform belongs to the
natriuretic peptide-sensitive mGCs. Additionally, CNP was able to reverse
the chloride inhibition of BTSM mGC activity, suggesting that this is a nov
el G protein-coupled GC-B receptor.