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Spring 2010
Effect of pH on disulfide bond reduction and enzyme activity
Effect of TCEP and temperature on circular dichroism spectrum (am class)
Effect of TCEP and temperature on circular dichroism spectrum (pm class)
Effect of reducing agents on thermal changes in Trp fluorescence in AP (am class)
Effect of reducing agents on thermal changes in Trp fluorescence in AP (pm class)
(These two groups also included temperature effects on free Trp fluorescence as well).
Two other groups performed limited proteolysis studies on the enzyme but did not complete until after the oral presentations were made.
Fall 2010
Analysis of Activity loss with AP purification
ANS and AP fluorescence
Fluorescence of free Trp and Tyr and Fast Red Dye Assay
Kinetic studies of temperature dependent loss of Trp Fluorescence of AP
CD temperature dependence rate of AP unfolding
CD 220 nm temperature dependence kinetics of AP
Effect of chelating agents on AP activity
Effect of EDTA on AP activity
Effects of high NaCl and MgCl2 on AP activity
Effects of pH on AP reduction by TCEP, DTT, and BME
Concentration dependency of TCEP inactivation of AP
Limited proteolysis of AP using TCEP and DTT
Spring 2011
CD spectral changes occurring upon thermal denaturation of AP: effects of SO4 vs. Cl-
ANS binding to AP during thermal denaturation
Effects of Salts and Chaperone-like proteins on thermal stability of AP
Fall 2011
Effects of Cationic Salts on AP Thermal Stability
EDTA and DTPA Chelation effects on AP (exp1)
edta_dtpa_inactivation of AP (exp 2)
Fluorescence changes using GdmHCl_am
Fluorescence changes using GdmHCL_pm
Limited Proteolysis_pm
Limited Proteolysis_am
Removal and Reintroduction of Zn and Mg
Tempdep of Ki and Km_am
Tempdep of Ki and Km_pm
Urea and GdmHCl effects seen in CD
urea_denaturation(exp 2) : these two are really only good for concentration and procedures, we saw no denaturation using fluorescence in either experiment, agreeing with the CD expt given above.
urea_denaturation(exp 1)
Biochemistry 463a
http://www.biochem.arizona.edu/classes/bioc463a/463a.html
Department of Biochemistry and Molecular Biophysics
The University of Arizona
jhazzard@email.arizona.edu
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Last revision Dec. 20 ,2001
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