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Reduced puromycin sensitivity of translocated polysomes after the addition of elongation factor 2 and non‐hydrolysable GTP analogues
Author(s) -
Nilsson Lars,
Nygård Odd
Publication year - 1992
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(92)80746-4
Subject(s) - puromycin , gtp' , elongation , polysome , elongation factor , chemistry , biochemistry , protein biosynthesis , microbiology and biotechnology , ribosome , biology , rna , enzyme , materials science , gene , metallurgy , ultimate tensile strength
Treatment of reticulocyte polysomes with elongation factor cEF‐2 and GTP led to an increased sensitivity or peptidyl‐tRNA for puromycin as a result of the translocation from the ribosomal A‐site to the P‐site, upon addition of an excess of the non‐hydrolysable GTP analogue, Guo PP (CH 2 ] P , the puromycin sensitivity decreased rapidly, The decrease in sensitivity required high concentrations of cEF‐2 with half maximal effect at an cEF‐2 concentration of around 1 μM. The data suggest either that peptidyl‐tRNA had re‐translocated back to the A‐site due to the higher affinity of cEF‐2 for the pre‐translocation than for the post‐translocation ribosome, or that the cEF‐2‐Guo PP [CH 2 ] P complex blocks the peptidyl‐transferase activity.