Abstract
The concept of using a covalently-linked chromophoric group to ‘report’ on the environment of an enzyme’s active site was first expounded by Burr and Koshland (1964). In the work reported here, we have used 3, 4-dihydro-3-methyl-6-nitro-2H-1,3-benzoxazin-2-one (DMNB) to provide a reporter group for examining the nature of the active site in cytosolic aldehyde dehydrogenase from sheep liver.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Preview
Unable to display preview. Download preview PDF.
Similar content being viewed by others
References
Abriola, D.P., MacKerell, A.D. and Pietruszko, R., 1990, Correlation of loss of activity of human aldehyde dehydrogenase with reaction of bromoacetophenone with glutamic acid-268 and cysteine-302 residues, Biochem. J., 266: 179–187.
Ambroziak, W., Kosley, L.L. and Pietruszko, R., 1989, Human aldehyde dehydrogenase: coenzyme binding studies, Biochemistry, 28: 5367–5373.
Blackwell, L.F., MacGibbon, A.K.H. and Buckley, P.D., 1989, Aldehyde dehydrogenases — kinetic characterisation, In Human Metabolism of Alcohol, vol. II (Crow, K.E. and Batt, R.D., eds), pp. 89–104, CRC Press, Boca Raton, Florida.
Blatter, E.E., Tasayco, M.L., Prestwich, G. and Pietruszko, R., 1990, Chemical modification of aldehyde dehydrogenase by a vinyl ketone analogue of an insect pheromone, Biochem. J., 272: 351–358.
Blatter, E.E., Abriola, D.P. and Pietruszko, R., 1992, Aldehyde dehydrogenase: covalent intermediate in aldehyde dehydrogenation and ester hydrolysis, Biochem. J., 282: 353–360.
Burr, M. and Koshland, D.E., 1964, Use of ‘reporter groups’ in structure-function studies of proteins, Proc. Natl. Acad. Sci. USA, 52: 1017–1024.
Deady, L.W., Buckley, P.D., Bennett, A.F. and Blackwell, L.F., 1985, Kinetics of inhibition and hysteresis of sheep liver cytoplasmic aldehyde dehydrogenase with glyoxylic acid: further evidence relating to the two-site model for aldehyde oxidation, Arch. Biochem. Biophys., 243: 586–597.
Ghenbot, G. and Weiner, H., 1992, Purification of liver aldehyde dehydrogenase by p-hydroxyacetophenone-Sepharose affinity matrix and the coelution of chloramphenicol acetyl transferase from the same matrix with recombinantly expressed aldehyde dehydrogenase, Protein Expression Purif., 3: 470–478.
Gutfreund, H., 1972, Enzymes: Physical Principles, pp. 68–72, Wiley, London.
Hempel, J.D. and Pietruszko, R., 1981, Selective chemical modification of human liver aldehyde dehydrogenases E, and E2 by iodoacetamide, J. Biol. Chem., 256:10889–10896.
Hempel, J.D., Nicholas, H. and Jörnvall, H., 1991, Thiol proteases and aldehyde dehydrogenases: evolution from a common thiolesterase precursor? Proteins: Struct., Funct. Genet., 11: 176–183.
Hempel, J.D., Nicholas, H. and Lindahl, R., 1993, Aldehyde dehydrogenases: widespread structural and functional diversity within a shared framework, Protein Science, 2: 1890–1900.
Kitson, T.M., 1976, The effect of some analogues of disulfiram on the aldehyde dehydrogenases of sheep liver, Biochem. J., 155:445–448.
Kitson, T.M., 1982, The activation of aldehyde dehydrogenase by diethylstilboestrol and 2,2′-dithiodipyridine, Biochem. J., 207: 81–89.
Kitson, T.M., 1989, Kinetics of p-nitrophenyl pivalate hydrolysis catalysed by cytoplasmic aldehyde dehydrogenase, Biochem. J., 257: 573–578.
Kitson, T.M. and Freeman, G.H., 1993, 3, 4-Dihydro-3-methyl-6-nitro-2H-l,3-benzoxazin-2-one, a reagent for labelling p-nitrophenyl esterases with a chromophoric reporter group — synthesis and reaction with chymotrypsin, Bioorg. Chem., 21: 354–365.
Kitson, T.M. and Kitson, K.E., 1994, Probing the active site of cytoplasmic aldehyde dehydrogenase with a chromophoric reporter group, Biochem. J., 300: 25–30.
Kitson, T.M., Hill, J.P. and Midwinter, G.G., 1991, Identification of a catalytically essential nucleophilic residue in sheep liver cytoplasmic aldehyde dehydrogenase, Biochem. J., 275: 207–210.
Leatherbarrow, R.J., 1987, Enzfitter Manual, Elsevier Science Publishers, Amsterdam.
McKenzie, H.A., 1969, pH and Buffers, In Data for Biochemical Research (Dawson, R.M.C., Elliott, D.C., Elliott, W.H. and Jones, K.M., eds), pp. 475–508, Oxford University Press, London.
Pietruszko, R., Abriola, D.P., Blatter, E.E. and Mukerjee, N., 1993, Aldehyde dehydrogenase: aldehyde dehydrogenation and ester hydrolysis, In Enzymology and Molecular Biology of Carbonyl Metabolism 4 (Weiner, H., Crabb, D.W. and Flynn, T.G., eds), pp. 221–232, Plenum Press, New York and London.
von Bahr-Lindström, H., Jeck, R., Woenckhaus, C., Sohn, S., Hempel, J. and Jörnvall, H., 1985, Characterisation of the coenzyme binding site of liver aldehyde dehydrogenase: differential reactivity of coenzyme analogues, Biochemistry, 24: 5847–5851.
Weiner, H., 1982, Aldehyde dehydrogenase, In Enzymology of Carbonyl Metabolism: Aldehyde Dehydrogenase and Aldo/Keto Reductase (Weiner, H. and Wermuth, B., eds), pp. 1–9, Alan R. Liss, New York.
Yoshida, A., Hsu, L.C. and Yanagawa, Y., 1993, Biological role of human cytosolic aldehyde dehydrogenase 1: hormonal response, retinal oxidation and implication in testicular féminisation, In Enzymology and Molecular Biology of Carbonyl Metabolism 4 (Weiner, H., Crabb, D.W. and Flynn, T.G., eds), pp. 37–44, Plenum Press, New York and London.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 1995 Springer Science+Business Media New York
About this chapter
Cite this chapter
Kitson, T.M., Kitson, K.E. (1995). Use of a Chromophoric Reporter Group to Probe the Active Site of Cytosolic Aldehyde Dehydrogenase. In: Weiner, H., Holmes, R.S., Wermuth, B. (eds) Enzymology and Molecular Biology of Carbonyl Metabolism 5. Advances in Experimental Medicine and Biology, vol 372. Springer, Boston, MA. https://doi.org/10.1007/978-1-4615-1965-2_5
Download citation
DOI: https://doi.org/10.1007/978-1-4615-1965-2_5
Publisher Name: Springer, Boston, MA
Print ISBN: 978-1-4613-5808-4
Online ISBN: 978-1-4615-1965-2
eBook Packages: Springer Book Archive