Characterization of guanylyl cyclase in purified myelin.
This study was undertaken to characterize the enzymatic properties of the particulate guanylyl cyclase previously shown to be present at a high level of activity in purified rat brain myelin. Significant activation was achieved by both Lubrol-PX and Triton X-100, the latter being somewhat more effective. A pH optimum of 7.8 was observed, compared to 7.4 for microsomes. Employing 1.2 mM GTP with 1% Triton X-100, linearity of response was observed up to 60 min and approximately 1.2 mg of myelin protein. Kinetic analysis revealed Km values of 0.258mM and 0.486mM for myelin and microsomes, respectively, similar values being obtained by Lineweaver-Burke analysis or Direct Linear Plot. Vmax values were 20 and 266 pmol/mg protein/min for myelin and microsomes, respectively. Washing of the myelin with 0.5 M NaCl or 0.1% Na taurocholate did not remove a significant amount of guanylyl cyclase activity, indicating the enzyme to be intrinsic to the myelin sheath.
Grabow
M
M
Department of Neurosciences, New Jersey Medical School, UMDNJ, Newark 07103, USA.
Chakraborty
G
G
Ledeen
R W
RW
D000818:Animals; D001933:Brain Stem / Q000201:enzymology*; D003902:Detergents; D004789:Enzyme Activation; D004795:Enzyme Stability; D006162:Guanylate Cyclase / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008297:Male; D008861:Microsomes / Q000201:enzymology; D009186:Myelin Sheath / Q000201:enzymology*; D017830:Octoxynol; D000077423:Polidocanol; D011092:Polyethylene Glycols; D051381:Rats; D017207:Rats, Sprague-Dawley
D003902:Detergents; D000077423:Polidocanol; D011092:Polyethylene Glycols; D017830:Octoxynol; D006162:Guanylate Cyclase