Biomaterial Database

Renaturation of lysozyme--temperature dependence of renaturation rate, renaturation yield, and aggregation: identification of hydrophobic folding intermediates. 1993

B Fischer, and I Sumner, and P Goodenough

AFRC Institute of Food Research, Department of Protein Engineering, Earley Gate, Reading, England.

Renaturation of denatured-reduced hen egg white lysozyme was analyzed at temperatures between 4 and 70 degrees C using the reduced/oxidized glutathione renaturation system. With an increase in temperature to 50 degrees C both renaturation rate constant and renaturation yield increased while formation of aggregates decreased. Denatured-reduced lysozyme and early folding intermediates were less stable against heat than native lysozyme at temperatures above 60 degrees C. Renaturation at 70 degrees C resulted in no reconstitution of lysozyme activity but the highest level of aggregation. Renaturation of denatured-reduced hen egg white lysozyme was further analyzed in the presence of the hydrophobicity-indicating fluorescence dye 1-anilinonaphalene-8-sulfonate at temperatures between 10 and 40 degrees C. The change in fluorescence intensity, the generation of enzyme activity, renaturation yield, and the formation of aggregates were studied. The results showed that early folding intermediates possess a strong hydrophobic nature. With an increase in temperature both the renaturation rate and the decay rate of hydrophobicity-mediated fluorescence increased. Consequently, with increasing temperature, accumulation of hydrophobic folding intermediates and formation of insoluble aggregates decreased, leading to an increase in the renaturation yield.

UI	MeSH Term	Description	Entries
D007700	Kinetics	The rate dynamics in chemical or physical systems.
D009113	Muramidase	A basic enzyme that is present in saliva, tears, egg white, and many animal fluids. It functions as an antibacterial agent. The enzyme catalyzes the hydrolysis of 1,4-beta-linkages between N-acetylmuramic acid and N-acetyl-D-glucosamine residues in peptidoglycan and between N-acetyl-D-glucosamine residues in chitodextrin. EC 3.2.1.17.	Lysozyme,Leftose,N-Acetylmuramide Glycanhydrolase,Glycanhydrolase, N-Acetylmuramide,N Acetylmuramide Glycanhydrolase
D011489	Protein Denaturation	Disruption of the non-covalent bonds and/or disulfide bonds responsible for maintaining the three-dimensional shape and activity of the native protein.	Denaturation, Protein,Denaturations, Protein,Protein Denaturations
D002645	Chickens	Common name for the species Gallus gallus, the domestic fowl, in the family Phasianidae, order GALLIFORMES. It is descended from the red jungle fowl of SOUTHEAST ASIA.	Gallus gallus,Gallus domesticus,Gallus gallus domesticus,Chicken
D006358	Hot Temperature	Presence of warmth or heat or a temperature notably higher than an accustomed norm.	Heat,Hot Temperatures,Temperature, Hot,Temperatures, Hot
D000818	Animals	Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA.	Animal,Metazoa,Animalia
D013816	Thermodynamics	A rigorously mathematical analysis of energy relationships (heat, work, temperature, and equilibrium). It describes systems whose states are determined by thermal parameters, such as temperature, in addition to mechanical and electromagnetic parameters. (From Hawley's Condensed Chemical Dictionary, 12th ed)	Thermodynamic
D017510	Protein Folding	Processes involved in the formation of TERTIARY PROTEIN STRUCTURE.	Protein Folding, Globular,Folding, Globular Protein,Folding, Protein,Foldings, Globular Protein,Foldings, Protein,Globular Protein Folding,Globular Protein Foldings,Protein Foldings,Protein Foldings, Globular
D046911	Macromolecular Substances	Compounds and molecular complexes that consist of very large numbers of atoms and are generally over 500 kDa in size. In biological systems macromolecular substances usually can be visualized using ELECTRON MICROSCOPY and are distinguished from ORGANELLES by the lack of a membrane structure.	Macromolecular Complexes,Macromolecular Compounds,Macromolecular Compounds and Complexes,Complexes, Macromolecular,Compounds, Macromolecular,Substances, Macromolecular