PMID: 9545533Apr 18, 1998Paper

Design of metal-binding green fluorescent protein variants

Biochimica Et Biophysica Acta
A BogdanovR Weissleder

Abstract

Diglycylcysteine motifs bind reduced oxo-compounds of technetium-99m, an important isotope in nuclear imaging. We suggested a system for detecting gene expression employing the effect of oxo[99mTc]technetate (Tc(V)O3+) transchelation and coordination with redox amino acid motifs. DNA fragments encoding diglycylcysteine (GGC) binding motifs were prepared by PCR and positioned downstream from the green fluorescent protein (GFP) cDNA insert. Using a Bluescript (+) vector with the fusion protein positioned under the control of a lac promoter, we obtained several E. coli clones expressing the following GFP fusion peptides: (1) GFP-P1 bearing a 'hydrophilic' C-terminal peptide (LEGGGCEGGC) containing two residues of glutamic acid and C-terminal cysteine (2) GFP-P2 carrying a 'hydrophobic' (LGGGGCGGGCGI) peptide (3) a control GFP fusion peptide with deleted C-terminal portion. Bacterial lysates obtained from the corresponding clones were tested for oxo[99mTc] technetate transchelation from a glucoheptonate complex. We found, using a solid phase assay, that radioactivity associated with protein lysates obtained from clones expressing GFP-P2 fusions were 3-4 fold higher than lysates prepared from a clone expressing a truncated GFP fusio...Continue Reading

References

Apr 1, 1995·Nuclear Medicine and Biology·H P VanbilloenA M Verbruggen
Jan 1, 1995·Annual Review of Microbiology·J C GloriosoD J Fink
Nov 1, 1995·Molecular Microbiology·C H ContagD A Benaron
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Citations

Feb 3, 2000·Annals of Nuclear Medicine·U Haberkorn
Aug 6, 2000·European Journal of Radiology·P WunderbaldingerR Weissleder
Nov 17, 2001·Seminars in Nuclear Medicine·P RayS S Gambhir
Apr 8, 2004·Cancer Biotherapy & Radiopharmaceuticals·Nico MertensChristophe Van de Wiele
Sep 9, 1999·Radiology·R Weissleder
Apr 27, 2001·Radiology·R Weissleder, U Mahmood
Jan 22, 2004·Seminars in Nuclear Medicine·Uwe HaberkornMichael Eisenhut
Jul 11, 2013·PloS One·Angel Goñi-MorenoFernando de la Cruz
Jan 29, 2003·Journal of Cellular Biochemistry. Supplement·Uwe Haberkorn, Annette Altmann
Apr 25, 2001·Gene Therapy·A BogdanovR Weissleder
Feb 21, 2006·FEMS Microbiology Letters·Rob Van HoudtChris W Michiels
Oct 24, 2006·FEMS Microbiology Letters·Morten GjermansenTim Tolker-Nielsen
Jan 20, 2007·FEMS Microbiology Letters·Laura Steindler, Vittorio Venturi
Aug 19, 2007·FEMS Microbiology Letters·Jana MüllerAlfred M Spormann
May 22, 2013·Proceedings of the National Academy of Sciences of the United States of America·Jeongjoon Choi, Eduardo A Groisman
Oct 24, 2003·Annals of Medicine·Uwe Haberkorn, Annette Altmann
Apr 15, 2008·Bioconjugate Chemistry·Zoia LevashovaFrancis G Blankenberg
Mar 12, 2003·Genes & Development·Tarik F Massoud, Sanjiv S Gambhir

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