Analysis of the T beta gamma-binding domain of MEKA/phosducin.

ABSTRACT

MEKA/phosducin, a 33 kDa phosphoprotein in the photoreceptor cell, associates with transducin beta gamma (T beta gamma) with its N-terminal domain (N-terminal 105 amino acids of MEKA), and translocates T beta gamma from the photoreceptor disc membrane to the soluble fraction. The present study further localized the T beta gamma-binding domain to aa 17-105 of MEKA, and showed that the activity of MEKA to translocate T beta gamma depends on the domain. A series of deletion mutant MEKA proteins were prepared to investigate the domain of MEKA which binds to and translocates T beta gamma. Both binding and translocation activities were not impaired by the deletion of the N-terminal 16 amino acids of MEKA, but completely abolished by further deletion to 42Val. Although anti-MEKA serum inhibited the T beta gamma-MEKA association, the antiserum absorbed with a recombinant peptide corresponding to aa 17-105 of MEKA did not, confirming that aa 17-105 of MEKA directly interacts with T beta gamma.