Lysozyme kinetic analysis

The effects of dimethyl sulphoxide, lithium bromide, guanidinium chloride, sodium dodecyl sulphate, and urea on lysozyme have been studied using Raman spectroscopy. The spectrum observed was found to depend on the denaturant used, suggesting there is not a unique denatured state for lysozyme. An analysis of the interaction of sodium dodecyl sulphate with lysozyme has been published. A kinetic study of the denaturation and subsequent reduction of disulphide bonds in lysozyme has been made using rapid ultrasonic absorption measurements. 

T4 lysozyme 33,497 helix stability of 528, 529 hydrophobic core stability of 533, 544 Tanford j8 value 544, 555, 578, 582-Temperature jump 137, 138, 541 protein folding 593 Terminal transferase 408,410 Ternary complex 120 Tertiary structure 22 Theorell-Chance mechanism 120 Thermodynamic cycles 125-131 acid denaturation 516,517 alchemical steps 129 double mutant cycles 129-131, 594 mutant cycles 129 specificity 381, 383 Thermolysin 22, 30,483-486 Thiamine pyrophosphate 62, 83 - 84 Thionesters 478 Thiol proteases 473,482 TNfn3 domain O-value analysis 594 folding kinetics 552 Torsion angle 16-18 Tbs-L-phenylalanine chloromethyl ketone (TPCK) 278, 475 Transaldolase 79 Tyransducin-o 315-317 Transit time 123-125 Transition state 47-49 definition 55... 

Dewey, T.G. (1994) Fractal analysis of proton exchange kinetics in lysozyme. Proc Natl Acad Sci USA, 91 (25), 12101-12104. 

Molecular mass determination by MS is also useful for analysis of substrate specificities and cleavage patterns of enzymes, including chitinases, chitosanases, lysozymes, and chitin deacety-lases, as summarized with a few representative examples in Table 11.4. Continuous infusion of reaction mixtures into an ESI mass spectrometer (so called real time monitoring) was nsed to analyze the hydrolysis of D (z = 4 - 6) by several chitosanases and some mutants (Dennhart et al. 2008). ESl-MS is also snitable for simnltaneous measurement of individual kinetic constants of enzymes in mixtures of substrates, as reported for a bacterial sulfotransferase with A (z = 2 - 5) (Pi and Leary 2004). 

The adsorption experiments with lysozyme indicate that the trough method may be a way of obtaining the adsorption kinetics of other globular proteins, including )S-lactoglobulin. This is necessary for the complete analysis of the response of an adsorbed film to a dilatational deformation. In order to use the method, however, reliable values of n and A area are required. 

A new technique for the rapid measurement of ultrasonic absorption with a sampling interval of 5 msec has been developed and applied to a kinetic study of denaturation and subsequent reduction of hen egg-white lysozyme. The rate constant of denaturation by guanidinium chloride obtained from the time variance of ultrasonic absorption agreed well with that from u.v. absorption and optical rotation. Analysis of the constants as functions of reagent concentrations indicated that the intermediate state between native and reduced state is not necessarily the completely denatured state. 

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