Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2007 May 1;364(1):92-4.
doi: 10.1016/j.ab.2007.01.019. Epub 2007 Jan 19.

Fluorescence-based analysis of aminoacyl- and peptidyl-tRNA by low-pH sodium dodecyl sulfate-polyacrylamide gel electrophoresis

Robert N Kirchdoerfer  1 , Joseph J-T Huang, Molly K Isola, Silvia Cavagnero
Affiliations

Fluorescence-based analysis of aminoacyl- and peptidyl-tRNA by low-pH sodium dodecyl sulfate-polyacrylamide gel electrophoresis

Robert N Kirchdoerfer et al. Anal Biochem. .
No abstract available

PubMed Disclaimer

Figures

Fig. 1
Fig. 1
Analysis of BODIPYFL-Met-tRNA fMet incubated under different conditions by low pH TA SDS-PAGE. BODIPYFL-Met-tRNAfMet samples were hydrolyzed in 0.1 M tris buffers, adjusted to pH values ranging from 6. to 9 by addition of aqueous HCl, at 37 °C for 1.5 hrs. Bands corresponding to BODIPYFL-Met-tRNAfMet and its ester bond hydrolysis products are enclosed in green and orange rectangles, respectively.
Fig. 2
Fig. 2
(A) Analysis of cell-free-generated peptidyl-tRNAs bearing N-terminal portions of the amino acid sequence of apoMb by low pH TA (Lanes 1, 2, 5 and 6) and reference pH TT (Lanes 3, 4, 7, and 8) SDS PAGE. All ribosome-associated nascent apoMb polypeptides were N-terminally labeled with BODIPYFL-Met-tRNAfMet. Bands corresponding to nascent polypeptides covalently linked to tRNA and tRNA-released by treatment with puromycin are enclosed in green and orange rectangles, respectively. (B) Diagram providing the quantitative extent of ester hydrolysis for peptidyl-tRNA run on the TA and TT gels in panel A (lanes 1, 3, 5, and 7). For each lane, the percent of ester hydrolysis was determined as the ratio of hydrolyzed product to total fluorescence intensity.
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Voigt J, Nagel K. Regulation of elongation factor g GTPase activity by the ribosomal state. J Biol Chem. 1993;268:100–106. - PubMed
    1. Plapp FV, Tilzer L, Hayes LC, Chiga M. A rapid method for isolation of peptidyl-tRNA from liver polysomes. Anal Biochem. 1975;64:170–176. - PubMed
    1. Kiely ML, McKnight GS, Schimke RT. Studies on attachment of carbohydrate to ovalbumin nascent chains in hen oviduct. J Biol Chem. 1976;251:5490–5495. - PubMed
    1. Kirk TZ, Evans JS, Veis A. Biosynthesis of type I procollagen. J Biol Chem. 1987;262:5540–5545. - PubMed
    1. Gite S, Mamaev S, Olejnik J, Rothschild K. Ultrasensitive fluorescence-based detection of nascent proteins in gels. Anal Biochem. 2000;279:218–225. - PubMed

Publication types

MeSH terms

LinkOut - more resources