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Effects Of Drying Methods And Additives On Structure And Function Of Actin: Mechanisms Of Dehydration-induced Damage And Its Inhibition.
S. D. Allison, T. Randolph, M. Manning, K. Middleton, A. Davis, J. Carpenter
Published 1998 · Medicine, Chemistry
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Limited stability impedes the development of industrial and pharmaceutical proteins. Dried formulations are theoretically more stable, but the drying process itself causes structural damage leading to loss of activity after rehydration. Lyophilization is the most common method used to dry proteins, but involves freezing and dehydration, which are both damaging to protein. We compared an air-drying method to freeze-drying to test the hypothesis that terminal dehydration is the critical stress leading to loss of activity. The secondary structure of air-dried and freeze-dried actin was analyzed by infrared spectroscopy and related to the level of activity recovered from the rehydrated samples. Actin dried by either method in the absence of stabilizers was highly unfolded and the capacity to polymerize was lost upon rehydration. The degree of unfolding was reduced by air-drying or freeze-drying actin with sucrose, and the level of activity recovered upon rehydration increased. The addition of dextran to sucrose improved the recovery of activity from freeze-dried, but not air-dried samples. Dextran alone failed to protect the structure and function of actin dried by either method, indicating that proteins are not protected from dehydration-induced damage by formation of a glassy matrix. In some cases, recovered activity did not correlate directly with the level of structural protection conferred by a particular additive. This result suggests that secondary structural protection during drying is a necessary but not sufficient condition for the recovery of activity from a dried protein after rehydration.
This paper references
Determination of protein secondary structure by Fourier transform infrared spectroscopy: a critical assessment.
W. Surewicz (1993)
Separation of freezing- and drying-induced denaturation of lyophilized proteins using stress-specific stabilization. II. Structural studies using infrared spectroscopy.
S. Prestrelski (1993)
Protein secondary structures in water from second-derivative amide I infrared spectra.
A. Dong (1990)
Surface-induced denaturation of proteins during freezing and its inhibition by surfactants.
B. Chang (1996)
Cryoprotective Effect of Saccharides on Denaturation of Catalase by Freeze-Drying
K. Tanaka (1991)
Infrared and laser-Raman spectroscopic studies of thermally-induced globular protein gels.
A. H. Clark (1981)
Calculation of protein conformation from circular dichroism.
J. Yang (1986)
Infrared methods for study of hemoglobin reactions and structures.
A. Dong (1994)
Optimization of Lyophilization Conditions for Recombinant Human Interleukin-2 by Dried-State Conformational Analysis Using Fourier-Transform Infrared Spectroscopy
S. Prestrelski (2004)
Stabilization of phosphofructokinase during air-drying with sugars and sugar/transition metal mixtures.
J. Carpenter (1987)
Nucleotide-free actin: stabilization by sucrose and nucleotide binding kinetics.
E. M. De La Cruz (1995)
Stabilization of phosphofructokinase with sugars during freeze-drying: characterization of enhanced protection in the presence of divalent cations.
J. Carpenter (1987)
A New Strategy for Enhancing the Stability of Lyophilized Protein: The Effect of the Reconstitution Medium on Keratinocyte Growth Factor
M. Zhang (2004)
Thermomechanical properties of small-carbohydrate–water glasses and ‘rubbers’. Kinetically metastable systems at sub-zero temperatures
H. Levine (1988)
An infrared spectroscopic study of the interactions of carbohydrates with dried proteins.
J. Carpenter (1989)
Optical rotatory dispersion and circular dichroic spectra of G-actin.
B. Nagy (1972)
Infrared spectroscopic studies of lyophilization- and temperature-induced protein aggregation.
A. Dong (1995)
Molten globule and protein folding.
O. Ptitsyn (1995)
The stabilization of proteins by sucrose.
J. Lee (1981)
Partition of Proteins in Liquid Polymer–Polymer Two-Phase Systems
P. Albertsson (1958)
Roger B. Gregory (1995)
Fluorimetry study of N-(1-pyrenyl)iodoacetamide-labelled F-actin. Local structural change of actin protomer both on polymerization and on binding of heavy meromyosin.
T. Kouyama (1981)
Proteins in frozen solutions: evidence of ice-induced partial unfolding.
G. Strambini (1996)
Separation of freezing- and drying-induced denaturation of lyophilized proteins using stress-specific stabilization. I. Enzyme activity and calorimetric studies.
J. Carpenter (1993)
Calorimetric study of the heat and cold denaturation of beta-lactoglobulin.
Y. Griko (1992)
Quantitation of the area of overlap between second-derivative amide I infrared spectra to determine the structural similarity of a protein in different states.
B. Kendrick (1996)
Effects of phase separating systems on lyophilized hemoglobin.
M. Heller (1996)
Control of aggregation in protein refolding: A variety of surfactants promote renaturation of carbonic anhydrase II
D. Wetlaufer (1995)
Chapter 18 Purification of Muscle Actin
J. D. Pardee (1982)
Dehydration-induced conformational transitions in proteins and their inhibition by stabilizers.
S. Prestrelski (1993)
Resolution-enhanced Fourier transform infrared spectroscopy of enzymes.
H. Susi (1986)
Stabilization of protein structure by sugars.
T. Arakawa (1982)
The frozen cellEdited by G. E. W. Wolstenholme and Maeve O'Connor
D. Pegg (1970)
Partitioning in aqueous two-phase systems: recent results.
H. Walter (1991)
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