Biotechnology Letters

, Volume 23, Issue 20, pp 1697–1702 | Cite as

Synchronous luminescence: a simple technique for the analysis of hydrolysis activity of the fragile histidine triad protein

  • Minoo Askari
  • Gordon Miller
  • Tuan Vo-Dinh


Human fragile histidine triad (FHIT) protein has dinucleoside 5′,5′′′-P1,Pn-polyphosphates hydrolysis activity, with AMP being one of the reaction products. Application of synchronous luminescence (SL) spectroscopy, in which both excitation and emission wavelengths are scanned simultaneously while a constant wavelength interval is maintained between them, was investigated for detection of the enzymatic activity of the FHIT protein. Ability of SL to identify reaction components, AMP production and its increase as a result of increase in substrate concentration and inhibition of the hydrolysis activity by ZnCl2 are demonstrated.

fragile histidine triad gene fluorescence spectroscopy hydrolysis synchronous luminescence spectroscopy 


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  1. Asensio A,Oaknin S,Rotllan P (1999) Fluorimetric detection of enzymatic activity associated with the human tumor suppressor Fhit protein. Biochim. Biophys. Acta 13: 396-400.Google Scholar
  2. Barnes LD,Robinson AK,Mumford CH,Garrison PN (1985) Assay of diadenosine tetraphosphate hydrolytic enzymes by boronate chromatography. Anal. Biochem. 144: 296-304.Google Scholar
  3. Brenner C,Pace H,Garrison P,Robinson A,Rosler A,Liu X,Blackburn G,Croce C,Huebner K,Barnes L (1997) Purification and crystallization of complexes modeling the active state of the fragile histidine triad protein. Protein Eng. 10: 1461-1463.Google Scholar
  4. Draganescu A,Hodawadekar S,Gee K,Brenner C (2000) FHITnucleotide specificity probed with novel fluorescent and fluorogenic substrates. J. Biol. Chem. 18: 4555-4560.Google Scholar
  5. Ji L,Fang B,Roth J (1999) A rapid, sensitive, and nonradioactive method for assay of FHIT Ap3A hydrolase activity by fluorescence thin-layer chromatographic image analysis. Anal. Biochem. 15: 114-116.Google Scholar
  6. Ohta M,Inoue H,Cotticelli M,Kastury K,Baffa R,Palazzo J,Siprashvili Z,Mori M,McCue P,Druck T,Croce C,Huebner K (1996) The FHIT gene, spanning the chromosome 3p14.2 fragile site and renal carcinoma-associated t(3;8) breakpoint, is abnormal in digestive tract cancers. Cell 84: 587-597.Google Scholar
  7. Sard L,Accornero P,Tornielli S,Delia D,Bunone G,Campiglio M,Colombo M,Gramegna M,Croce C,Pierotti M,Sozzi G (1999) The tumor-suppressor gene FHIT is involved in the regulation of apoptosis and in cell cycle control. Proc. Natl. Acad. Sci. USA 20: 8489-8492.Google Scholar
  8. Vo-Dinh T (1978) Synchronous luminescence for multi-component analysis. Anal. Chem. 50: 396.Google Scholar
  9. Vo-Dinh T (1981) Analytical measurements and instrumentation for process and pollution control. In: Cheremisinoff P,Perlis H, eds. Luminescence Spectrometry. New York: Ann Arbor Science Publishers, pp. 47-80.Google Scholar
  10. Vo-Dinh T,Gammage R,Martinez P (1981) Analysis of a workplace air particulate sample by synchronous luminescence and roomtemperature phosphorescence. Anal. Chem. 53: 253-258.Google Scholar
  11. Watts W,Isola N,Frazier D,Vo-Dinh T (1999) Differentiation of normal and neoplastic cells by synchronous fluorescence: rat liver epithelial and rat hepatoma cell models. Anal. Lett. 13: 2583-2594.Google Scholar

Copyright information

© Kluwer Academic Publishers 2001

Authors and Affiliations

  • Minoo Askari
    • 1
  • Gordon Miller
    • 1
  • Tuan Vo-Dinh
    • 1
  1. 1.Advanced Monitoring Development Group, Life Sciences DivisionOak Ridge National LaboratoryOak RidgeUSA

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