Interaction of mitochondrial elongation factors Tu.Ts with aminoacyl-tRNA.
The interaction between the bovine mitochondrial translational elongation factor Tu.Ts complex (EF-Tu.Tsmt) and aminoacyl-tRNA has been investigated using a nuclease protection assay and fluorescence enhancement of [AEDANS-s2C]Tyr-tRNA(Tyr). The equilibrium dissociation constant, Kd, for the EF-Tu.Tsmt:GTP:E. coli Phe-tRNA complex is approximately 50 nM. A similar binding constant (30 nM) is obtained using bovine mitochondrial Phe-tRNA. The equilibrium binding constant for the EF-Tu.Tsmt:GTP:yeast [AEDANS-s2C]Tyr-tRNA(Tyr) complex is approximately 4 nM when determined using the fluorescence enhancement assay.
Benkowski
L A
LA
University of North Carolina at Chapel Hill, Department of Chemistry 27599-3290, USA.
Takemoto
C
C
Ott
G
G
Beikman
M
M
Ueda
T
T
Watanabe
K
K
Sprinzl
M
M
Spremulli
L L
LL
D000818:Animals; D002417:Cattle; D004926:Escherichia coli / Q000378:metabolism; D005456:Fluorescent Dyes; D066298:In Vitro Techniques; D007700:Kinetics; D008928:Mitochondria / Q000378:metabolism; D009282:Naphthalenesulfonates; D010444:Peptide Elongation Factor Tu / Q000378:metabolism*; D010445:Peptide Elongation Factors / Q000378:metabolism*; D012346:RNA, Transfer, Amino Acyl / Q000378:metabolism*; D012360:RNA, Transfer, Phe / Q000378:metabolism; D012365:RNA, Transfer, Tyr / Q000378:metabolism; D012260:Ribonucleases / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000378:metabolism; D013050:Spectrometry, Fluorescence
D005456:Fluorescent Dyes; D009282:Naphthalenesulfonates; D010445:Peptide Elongation Factors; D012346:RNA, Transfer, Amino Acyl; D012360:RNA, Transfer, Phe; D012365:RNA, Transfer, Tyr; C023512:elongation factor Ts; C006421:1,5-I-AEDANS; D012260:Ribonucleases; D010444:Peptide Elongation Factor Tu