Comparison of particulate guanylate cyclase in cells with and without atrial natriuretic peptide receptor binding activity.
A line of kidney cells (PK1) which does not possess measurable ANP binding but has an active particulate guanylate cyclase has been identified. The physical characteristics of this enzyme were compared with those of particulate guanylate cyclase and ANP receptors isolated from rat lung. Although receptor and enzyme appear to reside on the same protein in the lung while the cyclase from PK1 cells does not possess ANP binding activity, these proteins exhibit identical physical characteristics. Guanylate cyclase from PK1 cells and rat lung and ANP receptor from lung co-eluted during gel filtration chromatography, with a Stokes radius of 6.1 nm. Also, these activities co-migrated through sucrose density gradients with S20,w values of 10.4 to 10.9. Using these parameters, a molecular weight of about 270 kD was estimated for all three activities. Furthermore, these enzyme activities exhibited similar mobilities in isoelectric focusing gels, with a pI of 6.1. Thus, although particulate guanylate cyclase from lung presumably possesses receptor binding activity, it is physically identical to a form of this enzyme associated with no measurable binding activity. Possible explanations for these observations are discussed.
Waldman
S A
SA
Department of Pharmacology, Stanford University School of Medicine, CA.
Leitman
D C
DC
Chang
L Y
LY
Murad
F
F
D000818:Animals; D009320:Atrial Natriuretic Factor / Q000378:metabolism*; D002460:Cell Line; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D006162:Guanylate Cyclase / Q000032:analysis* / Q000302:isolation & purification / Q000378:metabolism; D007668:Kidney / Q000032:analysis / Q000166:cytology / Q000201:enzymology*; D008168:Lung / Q000032:analysis / Q000166:cytology / Q000201:enzymology*; D051381:Rats; D017461:Receptors, Atrial Natriuretic Factor; D011956:Receptors, Cell Surface / Q000032:analysis* / Q000378:metabolism
D011956:Receptors, Cell Surface; D009320:Atrial Natriuretic Factor; D006162:Guanylate Cyclase; D017461:Receptors, Atrial Natriuretic Factor