Temperature adaptation of proteins: Engineering mesophilic-like activity and stability in a cold-adapted alpha-amylase

[en] Two multiple mutants of a psychrophilic alpha-amylase were produced, bearing five mutations (each introducing additional weak interactions found in pig pancreatic (alpha-amylase) with or without an extra disulfide bond specific to warm-blooded animals. Both multiple mutants display large modifications of stability and activity arising from synergic effects in comparison with single mutations. Newly introduced weak interactions and the disulfide bond confer mesophilic-like stability parameters, as shown by increases in the melting point t(m), in the calorimetric enthalpy DeltaH(cal) and in protection against heat inactivation, as well as by decreases in cooperativity and reversibility of unfolding. In addition, both kinetic and thermodynamic activation parameters of the catalyzed reaction are shifted close to the values of the porcine enzyme. This study confirms the central role of weak interactions in regulating the balance between stability and activity of an enzyme in order to adapt to the environmental temperature. (C) 2003 Elsevier Ltd. All rights reserved.

Disciplines :

Biochemistry, biophysics & molecular biology

Author, co-author :

D'Amico, Salvino ; Université de Liège - ULiège > GIGA-Research

Gerday, Charles ; Université de Liège - ULiège > Services généraux (Faculté des sciences) > Relations académiques et scientifiques (Sciences)

Feller, Georges ; Université de Liège - ULiège > Département des sciences de la vie > Labo de biochimie

Language :

English

Title :

Temperature adaptation of proteins: Engineering mesophilic-like activity and stability in a cold-adapted alpha-amylase

Publication date :

03 October 2003

Journal title :

Journal of Molecular Biology

ISSN :

0022-2836

eISSN :

1089-8638

Publisher :

Academic Press Ltd Elsevier Science Ltd, London, United Kingdom

Volume :

332

Issue :

Pages :

981-988

Peer reviewed :

Peer Reviewed verified by ORBi

Available on ORBi :

since 26 January 2010

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Bibliography

Margesin R., Feller G., Gerday C., Russell N. Cold-adapted microorganisms: adaptation strategies and biotechnological potential. Bitton G. The Encyclopedia of Environmental Microbiology. vol. 2:2002;871-885 Wiley, New York.
D'Amico S., Claverie P., Collins T., Georlette D., Gratia E., Hoyoux A., et al. Molecular basis of cold adaptation. Phil. Trans. Roy. Soc. ser. B. 357:2002;917-925.
Wintrode P.L., Arnold F.H. Temperature adaptation of enzymes: lessons from laboratory evolution. Advan. Protein Chem. 55:2000;161-225.
Feller G. Molecular adaptations to cold in psychrophilic enzymes. Cell. Mol. Life Sci. 60:2003;648-662.
Smalas A.O., Heimstad E.S., Hordvik A., Willassen N.P., Male R. Cold adaption of enzymes: structural comparison between salmon and bovine trypsins. Proteins: Struct. Funct. Genet. 20:1994;149-166.
Aghajari N., Feller G., Gerday C., Haser R. Crystal structures of the psychrophilic α-amylase from Alteromonas haloplanctis in its native form and complexed with an inhibitor. Protein Sci. 7:1998;564-572.
Aghajari N., Feller G., Gerday C., Haser R. Structures of the psychrophilic Alteromonas haloplanctis α-amylase give insights into cold adaptation at a molecular level. Structure. 6:1998;1503-1516.
Alvarez M., Zeelen J.P., Mainfroid V., Rentier-Delrue F., Martial J.A., Wyns L., et al. Triose-phosphate isomerase (TIM) of the psychrophilic bacterium Vibrio marinus. Kinetic and structural properties. J. Biol. Chem. 273:1998;2199-2206.
Russell R.J., Gerike U., Danson M.J., Hough D.W., Taylor G.L. Structural adaptations of the cold-active citrate synthase from an Antarctic bacterium. Structure. 6:1998;351-361.
Kim S.Y., Hwang K.Y., Kim S.H., Sung H.C., Han Y.S., Cho Y.J. Structural basis for cold adaptation. Sequence, biochemical properties, and crystal structure of malate dehydrogenase from a psychrophile Aquaspirillium arcticum. J. Biol. Chem. 274:1999;11761-11767.
de Backer M., McSweeney S., Rasmussen H.B., Riise B.W., Lindley P., Hough E. The 1.9 Å crystal structure of heat-labile shrimp alkaline phosphatase. J. Mol. Biol. 318:2002;1265-1274.
Toyota E., Ng K.K., Kuninaga S., Sekizaki H., Itoh K., Tanizawa K., James M.N. Crystal structure and nucleotide sequence of an anionic trypsin from chum salmon (Oncorhynchus keta) in comparison with Atlantic salmon (Salmo salar) and bovine trypsin. J. Mol. Biol. 324:2002;391-397.
Matsuura A., Yao M., Aizawa T., Koganesawa N., Masaki K., Miyazawa M., et al. Structural analysis of an insect lysozyme exhibiting catalytic efficiency at low temperatures. Biochemistry. 41:2002;12086-12092.
Aghajari N., Van Petegem F., Villeret V., Chessa J.P., Gerday C., Haser R., Van Beeumen J. Crystal structures of a psychrophilic metalloprotease reveal new insights into catalysis by cold-adapted proteases. Proteins: Struct. Funct. Genet. 50:2003;636-647.
Van Petegem F., Collins T., Meuwis M.A., Gerday C., Feller G., Van Beeumen J. The structure of a cold-adapted family 8 xylanase at 1.3 Å resolution. Structural adaptations to cold and investigation of the active site. J. Biol. Chem. 278:2003;7531-7539.
Vieille C., Zeikus G.J. Hyperthermophilic enzymes: sources, uses, and molecular mechanisms for thermostability. Microbiol. Mol. Biol. Rev. 65:2001;1-43.
D'Amico S., Marx J.C., Gerday C., Feller G. Activity-stability relationships in extremophilic enzymes. J. Biol. Chem. 278:2003;7891-7896.
Lonhienne T., Gerday C., Feller G. Psychrophilic enzymes: revisiting the thermodynamic parameters of activation may explain local flexibility. Biochim. Biophys. Acta. 1543:2000;1-10.
Collins T., Meuwis M.A., Gerday C., Feller G. Activity, stability and flexibility in glycosidases adapted to extreme thermal environments. J. Mol. Biol. 328:2003;419-428.
D'Amico S., Gerday C., Feller G. Structural similarities and evolutionary relationships in chloride-dependent alpha-amylases. Gene. 253:2000;95-105.
Janecek S. α-Amylase family: molecular biology and evolution. Prog. Biophys. Mol. Biol. 67:1997;67-97.
D'Amico S., Gerday C., Feller G. Structural determinants of cold adaptation and stability in a large protein. J. Biol. Chem. 276:2001; 25791-25796.
Qian M., Haser R., Buisson G., Duee E., Payan F. The active center of a mammalian α-amylase. Structure of the complex of a pancreatic α-amylase with a carbohydrate inhibitor refined to 2.2-Å resolution. Biochemistry. 33:1994;6284-6294.
D'Amico S., Gerday C., Feller G. Dual effects of an extra disulfide bond on the activity and stability of a cold-adapted α-amylase. J. Biol. Chem. 277:2002;46110-46115.
Feller G., le Bussy O., Houssier C., Gerday C. Structural and functional aspects of chloride binding to Alteromonas haloplanctis α-amylase. J. Biol. Chem. 271:1996;23836-23841.
Feller G., le Bussy O., Gerday C. Expression of psychrophilic genes in mesophilic hosts: assessment of the folding state of a recombinant α-amylase. Appl. Environ. Microbiol. 64:1998;1163-1165.
Feller G., d'Amico D., Gerday C. Thermodynamic stability of a cold-active α-amylase from the Antarctic bacterium Alteromonas haloplanctis. Biochemistry. 38:1999;4613-4619.
Kumar S., Tsai C.J., Nussinov R. Maximal stabilities of reversible two-state proteins. Biochemistry. 41:2002;5359-5374.
Privalov P.L., Medved L.V. Domains in the fibrinogen molecule. J. Mol. Biol. 159:1982;665-683.
Vogl T., Jatzke C., Hinz H.J., Benz J., Huber R. Thermodynamic stability of annexin V E17G: equilibrium parameters from an irreversible unfolding reaction. Biochemistry. 36:1997;1657-1668.
Low P.S., Bada J.L., Somero G.N. Temperature adaptation of enzymes: roles of the free energy, the enthalpy, and the entropy of activation. Proc. Natl Acad. Sci. USA. 70:1973;430-432.
Lejeune A., Vanhove M., Lamotte-Brasseur J., Pain R.H., Frere J.M., Matagne A. Quantitative analysis of the stabilization by substrate of Staphylococcus aureus PC1 beta-lactamase. Chem. Biol. 8:2001;831-842.
Gianese G., Bossa F., Pascarella S. Comparative structural analysis of psychrophilic and meso- and thermophilic enzymes. Proteins: Struct. Funct. Genet. 47:2002;236-249.
Chakravarty S., Varadarajan R. Elucidation of factors responsible for enhanced thermal stability of proteins: a structural genomics based study. Biochemistry. 41:2002;8152-8161.
Suhre K., Claverie J.M. Genomic correlates of hyperthermostability, an update. J. Biol. Chem. 278:2003;17198-17202.
Lonhienne T., Baise E., Feller G., Bouriotis V., Gerday C. Enzyme activity determination on macromolecular substrates by isothermal titration calorimetry: application to mesophilic and psychrophilic chitinases. Biochim. Biophys. Acta. 1545:2001;349-356.