Kinetics and Binding Sites for Interaction of the Prefoldin with a Group II Chaperonin

Prefoldin is a jellyfish-shaped hexameric co-chaperone of the group II chaperonins. It captures a protein folding intermediate and transfers it to a group II chaperonin for completion of folding. The manner in which prefoldin interacts with its substrates and cooperates with the chaperonin is poorly understood. In this study, we have examined the interaction between a prefoldin and a chaperonin from hyperthermophilic archaea by immunoprecipitation, single molecule observation, and surface plasmon resonance. We demonstrate that Pyrococcus prefoldin interacts most tightly with its cognate and in the we by that this interaction is in with between the We that and of the prefoldin for and for the interaction with a with and chaperonin prefoldin that in which a of prefoldin substrates to the Prefoldin is a jellyfish-shaped hexameric co-chaperone of the group II chaperonins. It captures a protein folding intermediate and transfers it to a group II chaperonin for completion of folding. The manner in which prefoldin interacts with its substrates and cooperates with the chaperonin is poorly understood. In this study, we have examined the interaction between a prefoldin and a chaperonin from hyperthermophilic archaea by immunoprecipitation, single molecule observation, and surface plasmon resonance. We demonstrate that Pyrococcus prefoldin interacts most tightly with its cognate and in the we by that this interaction is in with between the We that and of the prefoldin for and for the interaction with a with and chaperonin prefoldin that in which a of prefoldin substrates to the that for the and of the of to of that in to folding in manner group in and of and group II in archaea and in the of The group is a that is by the In the group II chaperonin in a of of and of a co-chaperone to The of the group II from the and the hyperthermophilic that the the of in the of a of the The chaperonin in the and to a of the of chaperonin chaperonin from prefoldin from prefoldin from from surface plasmon chaperonin from chaperonin from chaperonin chaperonin from prefoldin from prefoldin from from surface plasmon chaperonin from chaperonin from the group II is by the by the of group II with a Prefoldin to in the of and of the by in the to is in prefoldin and group II have in prefoldin of of and of and The of the prefoldin from a of It a in that its of a with from The of the and is that for the of prefoldin a to that of prefoldin and to a protein in the by its In the of the to with prefoldin in a that is is with the of the for and interaction with the of of prefoldin the interaction and between the chaperonin and prefoldin is understood. The a for of and have that the prefoldin from Pyrococcus cooperates with in the of the protein in In the study, we have examined the interaction between a prefoldin and a chaperonin from hyperthermophilic archaea by immunoprecipitation, single molecule observation, and surface plasmon and the and chaperonin of that the and of the for and for interaction with a the of the prefoldin and chaperonin that the with a and for and of the The for of the in this for the of and for a and for from from and from of and in and by from by from the the The to the of and that by a with from the and a of with a from the The for the the with and and and of by by a the The by in of most by for it by a with a of and a with by the of the and the of for The by a The and the of by from Pyrococcus and and and in prefoldin from in this is a with in the a in the and of and It of Prefoldin with The to the between the prefoldin and the chaperonin in the of The for the of the between and the The and of the The for the and the and for to the of The for of to of the and for to the and for to the by and in for the by and by of a with of by the of the that the is to the the of the with to the by a with with for with by for of with The and from the by The of by The of to and that of to and The the that of the the in from with of with for in The by with the and and a from a and for the of and the by with a with a The from the with by a to and for of for and from The of and the of surface plasmon with a the of to the and of protein in the of prefoldin in the surface to to the The and and the by with the of from the with to the The for to with the and for the and for the the the by the the is is the and is the of the by that for the of of from by the with a with of a to with of for from the of by the of of of in by the with a with the of of and and the in the a of for the of by the of by with by the of with the of Prefoldin with the of the interaction between prefoldin and its cognate chaperonin is a protein for to between the chaperonin and prefoldin from and in the of a with which and by In the of the protein interaction between and In a that a of by in the of with with for and the with in the of the of that the prefoldin interacts with the chaperonin and that the interaction is by the of of Prefoldin and to the interaction between the prefoldin and a chaperonin by the of a single The chaperonin from in this we its which to for with a of and protein and a a The of by the of and that of by the of of the to that a of the in the of with the the to the folding of of the between and and and from the in the and in the The which the by the in the The of and The of the of and the interaction between the prefoldin and a we and by surface plasmon a the a The interaction to the to a the of the of to the The of and a of the the with of between and by surface plasmon a to and and the a to and and the with of prefoldin and to the by and the the the with the and of the the of for and for which with and from the in The for the for and to The in by the between the and interaction and to the for the interaction between and The the from to by the the of the in which the interaction the for the is of a between the interaction is with and single molecule and between and by in a between and the interaction between prefoldin and a chaperonin we by surface plasmon the of to chaperonins. with the of the chaperonin to In a group chaperonin from interaction with the of for to with that of for the chaperonin of a interaction between and and that the of the interaction between prefoldin and chaperonin from a to and and the a and and the of the Prefoldin the of the of prefoldin that this substrates a by its of the of and prefoldin the of the hexameric to with by to the of the prefoldin it is the in this we a of and by to the of and to the of protein from and the of hexameric with a from the the to with the substrates the The of from the of the and and examined The to the of and to and the the and that the of from the of the of a the of the we that and of the for with this of to from the of the to for a prefoldin a in the to have of from the of prefoldin for the of from and of of prefoldin and of and and The from the and the with of from by the with a with with the of a to with folding by the with a in The with of of and and and with of a of of for the of from and of of in the of a of by the with a of of in the of a of by the with a of of of in a The of the to to The of the of the with a a from the the of prefoldin to the of of the with a of prefoldin to the of prefoldin to the chaperonin The to which of the the of prefoldin with the We examined the interaction of the with by surface plasmon the the the that of the of a prefoldin in the that this for the interaction with and of in the interaction with a and and the between and by the The of the in in to the of the to in a manner with the of to in the the for that of is the to for the interaction of with its to in a of with the of The of the to for chaperonin a to the chaperonin the a prefoldin the and in the of the and its interaction with the of prefoldin to the of the chaperonin we the of and chaperonin of the in the of for the chaperonin with the by that the chaperonin for the interaction between the and by the the by the by that of the in a this study, we have the interaction between the prefoldin and a chaperonin from hyperthermophilic archaea by immunoprecipitation, single molecule observation, and We between and by with In the single molecule a of chaperonin with prefoldin and prefoldin and to a chaperonin for folding to the In the interaction with the chaperonin the and of to a chaperonin in a manner to with of the between prefoldin and chaperonin of the to the and It is that prefoldin and chaperonin of prefoldin and of the it is that the interaction is most by the The from the with and It to the of the with the which is to by the of to a this study, we for a interaction between prefoldin and its cognate the between and for we between and in the we interaction between and by we have that the between and is in and in it that the to that protein from by and of for interaction with and chaperonins. In the in the to the of and to for the interaction of prefoldin with we that the of is for the its is the interaction with with the from to the for the chaperonin is to that of which with the of to a The between and chaperonin is the for chaperonin of which for and that of the We have of of from the prefoldin by chaperonins. and in The for the which with the of the with the is to between and in the interaction with a which is for the to the of the a and a protein the from the prefoldin and it is that the interaction between the prefoldin and the chaperonin is by the of a to in the between the and of the of a protein the for the interaction between the prefoldin and chaperonin examined in that for the and of the of to of that in to folding in manner group in and of and group II in archaea and in the of The group is a that is by the In the group II chaperonin in a of of and of a co-chaperone to The of the group II from the and the hyperthermophilic that the the of in the of a of the The chaperonin in the and to a of the of chaperonin chaperonin from prefoldin from prefoldin from from surface plasmon chaperonin from chaperonin from chaperonin chaperonin from prefoldin from prefoldin from from surface plasmon chaperonin from chaperonin from the group II is by the by the of The group II with a Prefoldin to in the of and of the by in the to is in prefoldin and group II have in prefoldin of of and of and The of the prefoldin from a of It a in that its of a with from The of the and is that for the of prefoldin a to that of prefoldin and to a protein in the by its In the of the to with prefoldin in a that is is with the of the for and interaction with the of of prefoldin In the interaction and between the chaperonin and prefoldin is understood. The a for of and We have that the prefoldin from Pyrococcus cooperates with in the of the protein in In the study, we have examined the interaction between a prefoldin and a chaperonin from hyperthermophilic archaea by immunoprecipitation, single molecule observation, and surface plasmon and the and chaperonin of that the and of the for and for interaction with a the of the prefoldin and chaperonin that the with a and for and of the The for of the in this for the of and for a and for from from and from of and in and by from by from the the The to the of and that by a with from the and a of with a from the The for the the with and and and of by by a the The by in of most by for it by a with a of and a with by the of the and the of for The by a The and the of by from Pyrococcus and and and in prefoldin from in this is a with in the a in the and of and It of Prefoldin with The to the between the prefoldin and the chaperonin in the of The for the of the between and the The and of the The for the and the and for to the of The for of to of the and for to the and for to the by and in for the by and by of a with of by the of the that the is to the the of the with to the by a with with for with by for of with The and from the by The of by The of to and that of to and The the that of the the in from with of with for in The by with the and and a from a and for the of and the by with a with a The from the with by a to and for of for and from The of and the of surface plasmon with a the of to the and of protein in the of prefoldin in the surface to to the The and and the by with the of from the with to the The for to with the and for the and for the the the by the the is is the and is the of the by that for the of of from by the with a with of a to with of for from the of by the of of of in by the with a with the of of and and the in the a of for the of by the of by with by the of with the and for and of the The for of the in this for the of and for a and for from from and from of and in and by from by from the the The to the of and that by a with from the and a of with a from the The for the the with and and and of by by a the The by in of most by for it by a with a of and a with by the of the and the of for The by a The and the of by from Pyrococcus and and and in prefoldin from The in this is a with in the a in the and of and It of Prefoldin with The to the between the prefoldin and the chaperonin in the of The for the of the between and the The and of the The for the and the and for to the of The for of to of the and for to the and for to the by and in for the by and by of a with of by the of the that the is to the the of the with to the by a with with for with by for of with The and from the by The of by The of to and that of to and The the that of the the in from with of with for in The by with the and and a from a and for the The of and the by with a with a The from the with by a to and for of for and from The of and the of surface plasmon with a the of to the and of protein in the of prefoldin in the surface to to the The and and the by with the of from the with to the The for to with the and for the and for the the the by the the is is the and is the of the by that for the of of from by the with a with of a to with of for from the of by the of of of in by the with a with the of of and and the in the a of for the of by the of by with by the of with the of Prefoldin with the of the interaction between prefoldin and its cognate chaperonin is a protein for to between the chaperonin and prefoldin from and in the of a with which and by In the of the protein interaction between and In a that a of by in the of with with for and the with in the of the of that the prefoldin interacts with the chaperonin and that the interaction is by the of of Prefoldin and to the interaction between the prefoldin and a chaperonin by the of a single The chaperonin from in this we its which to for with a of and protein and a a The of by the of and that of by the of of the to that a of the in the of with the the to the folding of of the of and the interaction between the prefoldin and a we and by surface plasmon a the a The interaction to the to a the of the of to the The of and a of the the with of between and by surface plasmon a to and and the a to and and the with of prefoldin and to the by and the the the with the and of the the of for and for which with and from the in The for the for and to The in by the between the and interaction and to the for the interaction between and The the from to by the the of the in which the interaction the for the is of a between the interaction is with and single molecule and between and by in a between and the interaction between prefoldin and a chaperonin we by surface plasmon the of to chaperonins. with the of the chaperonin to In a group chaperonin from interaction with the of for to with that of for the chaperonin of a interaction between and and that the of the interaction between prefoldin and chaperonin from a to and and the a and and the of the Prefoldin the of the of prefoldin that this substrates a by its of the of and prefoldin the of the hexameric to with by to the of the prefoldin it is the in this we a of and by to the of and to the of protein from and the of hexameric with a from the the to with the substrates the The of from the of the and and examined The to the of and to and the the and that the of from the of the of a the of the we that and of the for with this of to from the of the to for a prefoldin a in the to have of from the of prefoldin for the of from and of of prefoldin and of and and The from the and the with of from by the with a with with the of a to with folding by the with a in The with of of and and and with of a of of for the of from and of of in the of a of by the with a of of in the of a of by the with a of of of in a The of the to to The of the of the with a a from the the of prefoldin to the of of the with a of prefoldin to the of prefoldin to the chaperonin The to which of the the of prefoldin with the We examined the interaction of the with by surface plasmon the the the that of the of a prefoldin in the that this for the interaction with and of in the interaction with a and and the between and by the The of the in in to the of the to in a manner with the of to in the the for that of is the to for the interaction of with its to in a of with the of The of the to for chaperonin a to the chaperonin the a prefoldin the and in the of the and its interaction with the of prefoldin to the of the chaperonin we the of and chaperonin of the in the of for the chaperonin with the by that the chaperonin for the interaction between the and by the the by the by that of the in a of Prefoldin with the of the interaction between prefoldin and its cognate chaperonin is a protein for to between the chaperonin and prefoldin from and in the of a with which and by In the of the protein interaction between and In a that a of by in the of with with for and the with in the of the of that the prefoldin interacts with the chaperonin and that the interaction is by the of of Prefoldin and to the interaction between the prefoldin and a chaperonin by the of a single The chaperonin from in this we its which to for with a of and protein and a a The of by the of and that of by the of of the to that a of the in the of with the the to the folding of of the of and the interaction between the prefoldin and a we and by surface plasmon a the a The interaction to the to a the of the of to the The of and a of the the with The the the the with the and of the the of for and for which with and from the in The for the for and to The in by the between the and interaction and to the for the interaction between and The the from to by the the of the in which the interaction the for the is of a between the interaction is with and single molecule between and the interaction between prefoldin and a chaperonin we by surface plasmon the of to chaperonins. with the of the chaperonin to In a group chaperonin from interaction with the of for to with that of for the chaperonin of a interaction between and and that the The of the Prefoldin the of the of prefoldin that this substrates a by its of the of and prefoldin the of the hexameric to with by to the of the prefoldin it is the in this we a of and by to the of and to the of protein from and the of hexameric with a from the the to with the substrates the The of from the of the and and examined The to the of and to and the the and that the of from the of the of a the of the we that and of the for with this of to from the of the to for a prefoldin a in the to have The of the to to The of the of the with a a from the the of prefoldin to the of of the with a of prefoldin to the of prefoldin to the chaperonin The to which of the the of prefoldin with the We examined the interaction of the with by surface plasmon the the the that of the of a prefoldin in the that this for the interaction with and between and by the The of the in in to the of the to in a manner with the of to in the the for that of is the to for the interaction of with its to in a of with the of The of the to for chaperonin a to the chaperonin the a prefoldin the and in the of the and its interaction with the of prefoldin to the of the chaperonin we the of and chaperonin of the in the of for the chaperonin with the by that the chaperonin this study, we have the interaction between the prefoldin and a chaperonin from hyperthermophilic archaea by immunoprecipitation, single molecule observation, and We between and by with In the single molecule a of chaperonin with prefoldin and prefoldin and to a chaperonin for folding to the In the interaction with the chaperonin the and of to a chaperonin in a manner to with of the between prefoldin and chaperonin of the to the and It is that prefoldin and chaperonin of prefoldin and of the it is that the interaction is most by the The from the with and It to the of the with the which is to by the of to a this study, we for a interaction between prefoldin and its cognate the between and for we between and in the we interaction between and by we have that the between and is in and in it that the to that protein from by and of for interaction with and chaperonins. In the in the to the of and to for the interaction of prefoldin with we that the of is for the its is the interaction with with the from to the for the chaperonin is to that of which with the of to a The between and chaperonin is the for chaperonin of which for and that of the We have of of from the prefoldin by chaperonins. and in The for the which with the of the with the is to between and in the interaction with a which is for the to the of the a and a protein the from the prefoldin and it is that the interaction between the prefoldin and the chaperonin is by the of a to in the between the and of the of a protein the for the interaction between the prefoldin and chaperonin examined in In this study, we have the interaction between the prefoldin and a chaperonin from hyperthermophilic archaea by immunoprecipitation, single molecule observation, and We between and by with In the single molecule a of chaperonin with prefoldin and prefoldin and to a chaperonin for folding to the In the interaction with the chaperonin the and of to a chaperonin in a manner to with The of the between prefoldin and chaperonin of the to the and It is that prefoldin and chaperonin of prefoldin and of the it is that the interaction is most by the The from the with and It to the of the with the which is to by the of to a In this study, we for a interaction between prefoldin and its cognate the between and for we between and in the we interaction between and by we have that the between and is in and in it that the to that protein from by and of for interaction with and chaperonins. In the in the to the of and to for the interaction of prefoldin with we that the of is for the its is the interaction with with the from to the for the chaperonin is to that of which with the of to a The between and chaperonin is the for chaperonin of which for and that of the We have of of from the prefoldin by chaperonins. and in The for the which with the of the with the is to between and in the interaction with a which is for the to the of the a and a protein the from the prefoldin and it is that the interaction between the prefoldin and the chaperonin is by the of a to in the between the and of the of a protein the for the interaction between the prefoldin and chaperonin examined in We and for of We for in the of

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