CHB11-1-3 is a glycosylation mutant of Chinese hamster ovary (CHO) cells, isolated by screening mutagenized cells for those with decreased intracellular lysosomal enzyme activity (C. W. Hall et al. (1986) Mol. Cell. Biochem. 72, 35–45). CHB11-1-3 synthesizes the lipid polyprenol, the metabolic precursor of dolichol, rather than dolichol, indicating a defect in polyprenol reductase. This defect was demonstrated previously in Lec9 CHO mutants, and cell fusion experiments confirmed that CHB11-1-3 is a member of this complementation group. A revertant of CHB11-1-3, CHBREV, isolated for its ability to grow at 39°C, synthesizes dolichol at near-normal levels. CHBREV is probably a second-site revertant, because it synthesizes three to four times as much polyprenol as CHB11-1-3 and exhibits a similar elevation in the specific activity of cis -prenyl transferase. This higher activity appears to reflect an increase in enzyme molecules rather than the presence of an activator or absence of an inhibitor. These results suggest that CHB11-1-3 is a “ K m ” mutant, because synthesis of higher amounts of the substrate of polyprenol reductase obviates the defect.
Archives of Biochemistry and Biophysics – Elsevier
Published: Jul 1, 1997
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