The region of alpha-lactalbumin recognized by GroEL.
The binding constants between disulfide-intact or various disulfide-reduced bovine alpha-lactalbumins and an Escherichia coli chaperonin, GroEL, were determined by using the equilibrium dialysis method. The disulfide-intact and one-disulfide (Cys6-Cys120)-reduced alpha-lactalbumins were shown not to bind with GroEL both in the presence and absence of Ca2+. The two-disulfide (Cys6-Cys120 and Cys28-Cys111)-reduced alpha-lactalbumin, which has the native-like tertiary structure in its beta-domain region and an unfolded alpha-domain in the presence of Ca2+, showed considerable binding with GroEL. The binding free energy of the two-disulfide-reduced alpha-lactalbumin in the presence of Ca2+ is close to that of the molten globule state of disulfide-intact alpha-lactalbumin. This result suggests that GroEL binds to the unfolded alpha-domain of alpha-lactalbumin regardless of the conformation of the beta-domain. The fully disulfide-reduced and two-disulfide-reduced alpha-lactalbumins were found to bind more strongly with GroEL in the absence of Ca2+ than the two-disulfide-reduced alpha-lactalbumin in the presence of Ca2+, thus indicating that the unfolding of the beta-domain of alpha-lactalbumin leads to stronger interaction with GroEL.
Department of Bioengineering, Faculty of Engineering, Soka University, Hachioji, Tokyo, 192-8577, Japan.
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A
Shimizu
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Tanba
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Ogata
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Ikeguchi
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Sugai
D001667:Binding, Competitive; D018834:Chaperonin 60 / Q000378:metabolism*; D002942:Circular Dichroism; D003956:Dialysis; D004220:Disulfides; D004926:Escherichia coli / Q000378:metabolism; D007768:Lactalbumin / Q000737:chemistry / Q000378:metabolism*; D011485:Protein Binding; D011487:Protein Conformation
D018834:Chaperonin 60; D004220:Disulfides; D007768:Lactalbumin