Synthesis of Receptor Sites for Vitamin B12 in Euglena gracilis

In Euglena gracilis, vitamin B₁₂ uptake follows a biphasic patternconsisting of an initial rapid phase followed by a slower secondaryphase. Chase experiments showed that vitamin B₁₂ was tightlybound to its receptor-sites during either rapid or slow phaseof uptake. The slow phase was markedly inhibited when Euglenacells were preincubated with cycloheximide for 30 min at a concentrationof 100µg/ml. When the preincubation time was longer than30 min, a gradual inhibition of the rapid phase occurred andreached 84% after 4 h. This inhibitory effect of cycloheximideis reversible. On the other hand, tunicamycin at 1 µ/mlirreversibly inhibited the B₁₂ uptake suggesting that the receptor-sitefor B₁₂ is glycoprotein in nature. These results suggest thatthe rapid phase is also dependent on protein synthesis and representsB₁₂ binding on preexisting free receptors whereas the secondaryslow phase represents B_l2 binding on newly synthesized receptors.Both phases of uptake seem to be controled by the same receptor.The half-life of the free receptor is estimated to be 66 minwhereas the B₁₂ receptor complex is very stable. (Received October 3, 1988; Accepted February 15, 1989)