Activation of a chloroplast type of fructose bisphosphatase from Chlamydomonas reinhardtii by light-mediated agents.
A chloroplast type of fructose-1,6-bisphosphatase, a central regulatory enzyme of photosynthetic carbon metabolism, has been partially purified from Chlamydomonas reinhardtii. Unlike its counterpart from spinach chloroplasts, the algal FBPase showed a strict requirement for a dithiol reductant irrespective of Mg2+ concentration.  The enzymes from the two sources resembled each other immunologically, in subunit molecular mass and response to pH.  In the presence of dithiothreitol, the pH optimum for both the algal and spinach enzymes shifted from 8.5 to a more physiologic value of 8.0 as the Mg2+ concentration was increased from 1 to 16 mM.  At 1 mM Mg2+, a concentration estimated to be close to physiological, the Chlamydomonas FBPase was active only in the presence of reduced thioredoxin and was most active with Chlamydomonas thioredoxin f.  Under these conditions, the enzyme showed a pH optimum of 8.0.  The data suggest that the Chlamydomonas enzyme resembles its spinach counterpart in most respects, but it has a stricter requirement for reduction and less strict reductant specificity.   A comparison of the properties of the FBPases from Chlamydomonas and spinach will be helpful for elucidating the mechanism of the reductive activation of this enzyme.
Huppe
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HC
Division of Molecular Plant Biology, University of California, Berkeley 94720, USA.
Buchanan
B B
BB
D000818:Animals; D002244:Carbon / Q000378:metabolism; D016825:Chlamydomonas reinhardtii / Q000166:cytology / Q000201:enzymology* / Q000378:metabolism; D002736:Chloroplasts / Q000201:enzymology*; D004229:Dithiothreitol / Q000378:metabolism; D006597:Fructose-Bisphosphatase / Q000302:isolation & purification / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D008027:Light; D008274:Magnesium / Q000502:physiology; D010084:Oxidation-Reduction; D010788:Photosynthesis / Q000502:physiology / Q000528:radiation effects; D018724:Spinacia oleracea / Q000201:enzymology* / Q000378:metabolism; D013879:Thioredoxins / Q000378:metabolism
D013879:Thioredoxins; D002244:Carbon; D006597:Fructose-Bisphosphatase; D008274:Magnesium; D004229:Dithiothreitol