Nitric oxide activates the beta 2 subunit of soluble guanylyl cyclase in the absence of a second subunit.
Previously characterized mammalian soluble guanylyl cyclases form alpha/beta heterodimers that can be activated by the gaseous messenger, nitric oxide, and the novel guanylyl cyclase modulator YC-1. Four mammalian subunits have been cloned named alpha(1), beta(1), alpha(2), and beta(2). The alpha(1)/beta(1) and alpha(2)/beta(1) heterodimeric enzyme isoforms have been rigorously characterized. The role of the beta(2) subunit has remained elusive. Here we isolate a novel variant of this subunit and show that the beta(2) subunit does not need to form heterodimers for catalytic activity because enzyme activity can be measured when it is expressed alone in Sf9 cells. In analogy to the beta(3) subunit recently isolated from the insect Manduca sexta, activity was dependent on the presence of 4 mm free Mn(2+). The EC(50) values for the NO-donor diethylamine/NO were shifted to the left by 1 order of magnitude as compared with the alpha(1)/beta(1) heterodimeric form. In the presence of the detergent Tween, NO sensitivity of beta(2) was abolished, but the enzyme could be activated by protoporphyrin IX, indicating removal of a prosthetic heme group and exchange for the heme precursor. We suggest that the beta(2) subunit is the first mammalian NO-sensitive guanylyl cyclase lacking a heterodimeric structure.
Koglin
M
M
Institut für Experimentelle und Klinische Pharmakologie, Universität Hamburg, Martinistrasse 52, D-20251 Hamburg, Germany.
Vehse
K
K
Budaeus
L
L
Scholz
H
H
Behrends
S
S
D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D019281:Dimerization; D004789:Enzyme Activation; D006162:Guanylate Cyclase / Q000737:chemistry / Q000378:metabolism*; D008345:Manganese / Q000494:pharmacology; D008969:Molecular Sequence Data; D009569:Nitric Oxide / Q000502:physiology*; D021122:Protein Subunits; D051381:Rats; D018411:Spodoptera
D021122:Protein Subunits; D009569:Nitric Oxide; D008345:Manganese; D006162:Guanylate Cyclase