Thermodynamics of folding, stabilization, and binding in an engineered protein--protein complex.

Härd, Torleif, 1959 (author): Gothenburg University,Göteborgs universitet,Institutionen för anatomi och cellbiologi,Institute of Anatomy and Cell Biology,Göteborgs Universitet

(creator_code:org_t)

2004-08-20
2004
English.
In: Journal of the American Chemical Society. - : American Chemical Society (ACS). - 0002-7863 .- 1520-5126. ; 126:36, s. 11220-30

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Abstract Subject headings

We analyzed the thermodynamics of a complex protein-protein binding interaction using the (engineered) Z(SPA)(-)(1) affibody and it's Z domain binding partner as a model. Free Z(SPA)(-)(1) exists in an equilibrium between a molten-globule-like (MG) state and a completely unfolded state, wheras a well-ordered structure is observed in the Z:Z(SPA)(-)(1) complex. The thermodynamics of the MG state unfolding equilibrium can be separated from the thermodynamics of binding and stabilization by combined analysis of isothermal titration calorimetry data and a separate van't Hoff analysis of thermal unfolding. We find that (i) the unfolding equilibrium of free Z(SPA)(-)(1) has only a small influence on effective binding affinity, that (ii) the Z:Z(SPA)(-)(1) interface is inconspicuous and structure-based energetics calculations suggest that it should be capable of supporting strong binding, but that (iii) the conformational stabilization of the MG state to a well-ordered structure in the Z:Z(SPA)(-)(1) complex is associated with a large change in conformational entropy that opposes binding.

TEKNIK OCH TEKNOLOGIER -- Industriell bioteknik -- Annan industriell bioteknik (hsv//swe)
ENGINEERING AND TECHNOLOGY -- Industrial Biotechnology -- Other Industrial Biotechnology (hsv//eng)

By the author/editor: Dincbas-Renqvist ...; Lendel, Christof ...; Dogan, Jakob; Wahlberg, Elisab ...; Härd, Torleif, 1 ...

About the subject

ENGINEERING AND TECHNOLOGY: ENGINEERING AND ...; and Industrial Biote ...; and Other Industrial ...

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