Characterization of the Unfolding Process of Lipocalin-type Prostaglandin D Synthase

Inui, Takashi; Ohkubo, Tadayasu; Emi, Maiko; Irikura, Daisuke; Hayaishi, Osamu; Urade, Yoshihiro

doi:10.1074/jbc.m209934200

Cited by 47 publications

(48 citation statements)

References 42 publications

(80 reference statements)

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“…This interpretation is qualitatively in agreement with that urea generally has a larger effect on the unfolding rate constant compared to the refolding rate-constant in protein folding studies (30). Urea activation of other enzymes has also been observed-including dihydrofolate reductase (13,14), prostaglandin D synthase (16,17), and biliverdin-Ixa (18). Possibly, the increased activities in these other enzymes are also mediated by a redistribution of existing structural states.…”

Section: Discussionsupporting

confidence: 85%

“…It has been observed that common denaturants like urea can increase activities of certain enzymes, including dihydrofolate reductase (13,14), Adk (15), prostaglandin D synthase (16,17), and biliverdin-Ixa (18). Various mechanisms for these increases have been proposed, for example removal of inhibition in biliverdin-Ixa (18) and structural reorganization in prostaglandin D synthase (16).…”

Section: Introductionmentioning

confidence: 99%

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Urea-Dependent Adenylate Kinase Activation following Redistribution of Structural States

Rogne

Wolf‐Watz

2016

Biophysical Journal

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Proteins are often functionally dependent on conformational changes that allow them to sample structural states that are sparsely populated in the absence of a substrate or binding partner. The distribution of such structural microstates is governed by their relative stability, and the kinetics of their interconversion is governed by the magnitude of associated activation barriers. Here, we have explored the interplay among structure, stability, and function of a selected enzyme, adenylate kinase (Adk), by monitoring changes in its enzymatic activity in response to additions of urea. For this purpose we used a 31 P NMR assay that was found useful for heterogeneous sample compositions such as presence of urea. It was found that Adk is activated at low urea concentrations whereas higher urea concentrations unfolds and thereby deactivates the enzyme. From a quantitative analysis of chemical shifts, it was found that urea redistributes preexisting structural microstates, stabilizing a substrate-bound open state at the expense of a substrate-bound closed state. Adk is rate-limited by slow opening of substrate binding domains and the urea-dependent redistribution of structural states is consistent with a model where the increased activity results from an increased rate-constant for domain opening. In addition, we also detected a strong correlation between the catalytic free energy and free energy of substrate (ATP) binding, which is also consistent with the catalytic model for Adk. From a general perspective, it appears that urea can be used to modulate conformational equilibria of folded proteins toward more expanded states for cases where a sizeable difference in solvent-accessible surface area exists between the states involved. This effect complements the action of osmolytes, such as trimethylamine N-oxide, that favor more compact protein states.

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Section: Discussionsupporting

confidence: 85%

Section: Introductionmentioning

confidence: 99%

Urea-Dependent Adenylate Kinase Activation following Redistribution of Structural States

Rogne

Wolf‐Watz

2016

Biophysical Journal

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“…A␤ (1-16), A␤ (25)(26)(27)(28)(29)(30)(31)(32)(33)(34)(35), A␤ (1-40), A␤ (1-42) (Peptide Institute, Osaka, Japan), and A␤ (1-28) (AnaSpec, San Jose, CA) were dissolved in 0.02% ammonia solution at 200 M. The A␤ fibril seed solution was prepared by incubation of 50 M A␤ peptides in 50 mM sodium phosphate (pH 7.5) and 100 mM NaCl for 7 days at 37°C.…”

Section: Methodsmentioning

confidence: 99%

“…A␤ peptides (50 M) were incubated at 37°C in the absence or presence of human L-PGDS/␤-trace purified from human CSF (34), recombinant human L-PGDS expressed in E. coli (35), or human CSF in 50 mM sodium phosphate (pH 7.5) and 100 mM NaCl. The seed-dependent and spontaneous A␤ aggregations were monitored with or without addition of A␤ seed (final concentration 10 g/ml) in a Hitachi F-4500 f luorescence spectrophotometer (Hitachi Software Engineering, Yokohama, Japan) at excitation and emission wavelengths of 446 and 490 nm, respectively, after the components had been mixed with a 200-fold volume of 50 mM glycine-NaOH (pH 8.5) containing 5 M ThT (Wako Pure Chemicals, Osaka, Japan) as described earlier (36).…”

mentioning

confidence: 99%

Lipocalin-type prostaglandin D synthase/β-trace is a major amyloid β-chaperone in human cerebrospinal fluid

Kanekiyo¹,

Ban

Aritake

et al. 2007

Proc. Natl. Acad. Sci. U.S.A.

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137

156

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“…Enzyme Assay-The activities of H-PGDS (15), L-PGDS (21), and microsomal PGE synthase (m-PGES, (22) were measured with 40 M [1-…”

Section: Expression and Purification Of Recombinant H-pgds-human H-pgdsmentioning

confidence: 99%

Structural and Functional Characterization of HQL-79, an Orally Selective Inhibitor of Human Hematopoietic Prostaglandin D Synthase

Aritake

Kado

Inoue

et al. 2006

Journal of Biological Chemistry

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104

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Characterization of the Unfolding Process of Lipocalin-type Prostaglandin D Synthase

Cited by 47 publications

References 42 publications

Urea-Dependent Adenylate Kinase Activation following Redistribution of Structural States

Urea-Dependent Adenylate Kinase Activation following Redistribution of Structural States

Lipocalin-type prostaglandin D synthase/β-trace is a major amyloid β-chaperone in human cerebrospinal fluid

Structural and Functional Characterization of HQL-79, an Orally Selective Inhibitor of Human Hematopoietic Prostaglandin D Synthase

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