Backbone dynamics of the N-terminal domain in E. coli DnaJ determined by 15N- and 13CO-relaxation measurements

Kai Huang, Ranajeet Ghose, John M. Flanagan, James H. Prestegard

Research output: Contribution to journalArticle

23 Citations (Scopus)

Abstract

The backbone dynamics of the N-terminal domain of the chaperone protein Escherichia coli DnaJ have been investigated using steady-state 1H-15N NOEs, 15N T1, T2, and T(1ρ) relaxation times, steady-state 13C(α- 13)CO NOEs, and 13CO T1 relaxation times. Two recombinant constructs of the N-terminal domain of DnaJ have been studied. One, DnaJ(1-78), contains the most conserved 'J-domain' of DnaJ, and the other, DnaJ(1-104), includes a glycine/phenylalanine rich region ('G/F' region) in addition to the 'J- domain'. DnaJ(1-78) is not capable of stimulating ATP hydrolysis by DhaK, despite the fact that all currently identified sites responsible for DnaJ- DnaK interaction are located in this region. DnaJ(1 104), on the other hand, retains nearly the full ATPase stimulatory activity of full length DnaJ. Recently, a structural analysis of these two molecules was presented in an effort to elucidate the origin of their functional differences [Huang, K., Flanagan, J. M., and Prestegard, J. H. (1999) Protein Science 8, 203-214]. Herein, an analysis of dynamic properties is presented in a similar effort. A generalized model-free approach with a full treatment of the anisotropic overall rotation of the proteins is used in the analysis of measured relaxation parameters. Our results show that internal motions on pico- to nanosecond time scales in the backbone of DnaJ(1-78) are reduced on the inclusion of the 'G/F' region, while conformational exchange on micro- to millisecond time scales increases. We speculate that the enhanced flexibility of residues on the slow time scale upon the inclusion of the 'G/F' region could be relevant to the ATPase stimulatory activity of DnaJ if an 'induced- fit' mechanism applies to DnaJ-DnaK interactions.

Original languageEnglish (US)
Pages (from-to)10567-10577
Number of pages11
JournalBiochemistry
Volume38
Issue number32
DOIs
StatePublished - Aug 10 1999

Fingerprint

F region
Escherichia coli
Relaxation time
Adenosine Triphosphatases
Escherichia coli Proteins
Carbon Monoxide
Phenylalanine
Structural analysis
Glycine
Hydrolysis
Proteins
Adenosine Triphosphate
Molecules

All Science Journal Classification (ASJC) codes

  • Biochemistry

Cite this

Huang, Kai ; Ghose, Ranajeet ; Flanagan, John M. ; Prestegard, James H. / Backbone dynamics of the N-terminal domain in E. coli DnaJ determined by 15N- and 13CO-relaxation measurements. In: Biochemistry. 1999 ; Vol. 38, No. 32. pp. 10567-10577.
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abstract = "The backbone dynamics of the N-terminal domain of the chaperone protein Escherichia coli DnaJ have been investigated using steady-state 1H-15N NOEs, 15N T1, T2, and T(1ρ) relaxation times, steady-state 13C(α- 13)CO NOEs, and 13CO T1 relaxation times. Two recombinant constructs of the N-terminal domain of DnaJ have been studied. One, DnaJ(1-78), contains the most conserved 'J-domain' of DnaJ, and the other, DnaJ(1-104), includes a glycine/phenylalanine rich region ('G/F' region) in addition to the 'J- domain'. DnaJ(1-78) is not capable of stimulating ATP hydrolysis by DhaK, despite the fact that all currently identified sites responsible for DnaJ- DnaK interaction are located in this region. DnaJ(1 104), on the other hand, retains nearly the full ATPase stimulatory activity of full length DnaJ. Recently, a structural analysis of these two molecules was presented in an effort to elucidate the origin of their functional differences [Huang, K., Flanagan, J. M., and Prestegard, J. H. (1999) Protein Science 8, 203-214]. Herein, an analysis of dynamic properties is presented in a similar effort. A generalized model-free approach with a full treatment of the anisotropic overall rotation of the proteins is used in the analysis of measured relaxation parameters. Our results show that internal motions on pico- to nanosecond time scales in the backbone of DnaJ(1-78) are reduced on the inclusion of the 'G/F' region, while conformational exchange on micro- to millisecond time scales increases. We speculate that the enhanced flexibility of residues on the slow time scale upon the inclusion of the 'G/F' region could be relevant to the ATPase stimulatory activity of DnaJ if an 'induced- fit' mechanism applies to DnaJ-DnaK interactions.",
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Backbone dynamics of the N-terminal domain in E. coli DnaJ determined by 15N- and 13CO-relaxation measurements. / Huang, Kai; Ghose, Ranajeet; Flanagan, John M.; Prestegard, James H.

In: Biochemistry, Vol. 38, No. 32, 10.08.1999, p. 10567-10577.

Research output: Contribution to journalArticle

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T1 - Backbone dynamics of the N-terminal domain in E. coli DnaJ determined by 15N- and 13CO-relaxation measurements

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