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The use of methods of Directed Evolution allow researchers to improve
and "direct" such important biotechnological processes
as biocatalysis. In addition, Directed Evolution allows scientists
to address fundamental questions of adaptation, enzyme function and
evolution. Important findings of mechanisms of antibiotic resistance
were made using methods of directed evolution.
Professor Salerno is combining methods of Directed Evolution with
Bioinformatics. He designs new algorithms and methods that will improve
creation of chimeric genes and allow synthesis of new proteins. His
work on the creation of novel primer design strategies results in
significant enhancement of the technique of PCR and allows to dramatically
augment the efficiency of the creation of synthetic genes.
Main publications in Directed Evolution:
• Book: Arnold F.H. (Ed):
Evolutionary Protein Design. New York: Academic Press; 2001. [Advances
in Protein Chemistry vol. 55.]
• Journal article: E.T.
Farinas, T. Bulter and F. H. Arnold, Directed enzyme evolution,
Current Opinion in Biotechnology 2001, 12:545–551
Online resources in Directed Evolution:
• Introductory
article by F. Arnold
Prof. Salerno's publications
Journal articles:
• J.C. Salerno, E. Erdogan,
and M.H. Hanna, Chimera production for protein design and directed
evolution, in submission
• Salerno,J.C. et al (1997)
An autoinhibitory control element defines calcium regulated isoforms
of NO synthase J. Biol. Chem. 272, 29769-29777
• Salerno, J. C., Martasek,
P., Williams, R. F., and Masters, B. S. S. (1997) Substrate And Substrate
Analog Binding To Endothelial Nitric Oxide Synthase - Electron Paramagnetic
Resonance As An Isoform-Specific Probe Of The Binding Mode Of Substrate
Analogs. Biochemistry 36: 11821 - 11827.
• Migita, C. T., Salerno,
J. C., Masters, B. S. S., Martasek, P. Mcmillan, K. and Ikedasaito,
M. (1997) Substrate Binding-Induced Changes In The Epr Spectra Of
The Ferrous Nitric Oxide Complexes Of Neuronal Nitric Oxide Synthase.
Biochemistry 36: 10987 -10992.
• Zehr, J. P., Harris,
D., Dominic, B., and Salerno, J. (1997) Structural Analysis Of The
Trichodesmium Nitrogenase Iron Protein - Implications For Aerobic
Nitrogen Fixation Activity. FEMS Microbiology Letters 153: 303 -
309.
• Martasek P, Miller RT,
Liu Q, Roman LJ, Salerno JC, Migita CT, Raman CS, Gross SS, Ikeda-Saito
M, (1998) The Cys331A mutant of neuronal nitric oxide synthase is
defective in arginine binding J. Biol. Chem. 273(52), 34799-34805
• Martasek, P., Roman,
L.J., Raman, C.S., Liu, Q., Salerno, J.C., Ikeda-Saito, M., Gross,
S.S., and Masters, B.S.S. (1998) The Cys331A mutant of nNOS is defective
in arginine binding Portland Press Proceedings: Biology of Nitric
Oxide, vol 6
• Hunter, D.J., Salerno,
J.C. and Ingledew, W.J. (1998) Angular dependence of electron paramagnetic
resonances of an azide-NO complex of cytochrome c oxidase: orientation
of the haem-copper axis in cytochrome aa3 from ox heart. B.B.A. 1364,
55-62
• Hastings SF, Kaysser,
Jiang F., SalernoJ.C. , Gennis R.B., Ingledew WJ (1998) Identification
of a stable semiquinone intermediate in the purified and membrane
bound ubiquinol oxidase-cytochrome bd from Escherichia coli. Eur
J Biochem 255, 317-2
• Masters, B.S.S., Miller,
R.T., Roman, L.J., Nishmura, J.S., Panda, S., Harris, D.E., Horowitz,
P., Shea, T.M., Zhang, J., Kim, J.-J., and Salerno, J.C. ( 1999)
Structural Aspects of the Flavoprotein Domains of Isoforms of Nitric
Oxide Synthase in: Flavins and Flavoproteins 1999: Proc. 13th Int.
Symp., Konstanz, Germany; Ghisla S, Kroneck P, Macheroux P, Sund
H, Editors; Agency for Scientific Publ., Berlin
• Hunter, D. J., Oganesyan,
V. S. Salerno, J. C. Butler, C. S. Ingledew, W. J. Thomson, A. J.
(2000). Angular dependence of perpendicular and parallel mode electron
paramagnetic resonance of oxidized beef heart cytochrome c oxidase.
Biophys J 78: 439-450.
• Smith, S.M.E., Sham,
C., Roman, L. , Martasek, P., and Salerno, J.C. (2001) Titration
of low Kd binding sites: binding of arginine analogs to nitric oxide
synthases Nitric Oxide Journal 5(5), 442-452
• Ingledew, W.J., Salerno,
J.C., Smith, SM.E. and Rich, P. (2002) Neuronal nitric oxide synthase
ligand and protein vibrations at the substrate binding site. A study
by FTIR. Biochemistry.;41(26):8377-84.
• Ghosh, D.K. and Salerno
J.C. (2003) Nitric oxide synthases: domain structure and alignment
in enzyme function and control. Front Biosci. 8: 193-209.
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