Kinetics and energetics of subunit dissociation/unfolding of TIM: the importance of oligomerization for conformational persistence and chemical stability of proteins.


Abstract

Kinetics of unfolding and refolding of rabbit muscle triosephosphate isomerase (TIM) were measured as a function of guanidine hydrochloride (GdnHCl) concentration. From the rate constants of these processes, the activation free-energy barriers (delta G++) were calculated using the Arrhenius equation. Assuming a linear dependence of delta G++ on the concentration of GdnHCl, activation energies in the absence of GdnHCl were estimated. The Gibbs free-energy change of dissociation/unfolding (delta G) was determined from GdnHCl unfolding curves in equilibrium. Using these data and the literature value for the bimolecular association rate constant of folded TIM monomers [Zabori, S., Rudolph, R., and Jaenicke, R. (1980) Z. Naturforsch. 35C, 999-1004], a model was developed that fully describes both kinetics and energetics of subunit dissociation/unfolding of TIM. Unfolded TIM monomers are susceptible to proteolytic digestion and thiol oxidation, while native TIM is resistant to both. The present model explains how the dimeric nature of TIM decreases the frequency of subunit unfolding by several orders of magnitude, thus increasing the chemical stability of the protein. Furthermore, the model also explains the recently demonstrated persistence (on a time scale of hours to days) of conformational heterogeneity of native TIM dimers [Rietveld, A. W. M., and Ferreira, S. T. (1996) Biochemistry 35, 7743-7751]. Again, it appears that the dimeric nature of TIM is essential for this behavior. Study holds ProTherm entries: 9477, 9478 Extra Details: dithiothreitol(1 mM) was added in the experiment triosephosphate isomerase; Arrhenius equation; activation energies;,chemical stability; conformational heterogeneity

Submission Details

ID: prRXvp6E4

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:38 p.m.

Version: 1

Publication Details
Rietveld AW;Ferreira ST,Biochemistry (1998) Kinetics and energetics of subunit dissociation/unfolding of TIM: the importance of oligomerization for conformational persistence and chemical stability of proteins. PMID:9454583
Additional Information

Structure view and single mutant data analysis

Study data

No weblogo for data of varying length.
Colors: D E R H K S T N Q A V I L M F Y W C G P
 

Data Distribution

Studies with similar sequences (approximate matches)

Correlation with other assays (exact sequence matches)


Relevant PDB Entries

Structure ID Release Date Resolution Structure Title
1R2R 2003-09-29T00:00:00+0000 1.5 CRYSTAL STRUCTURE OF RABBIT MUSCLE TRIOSEPHOSPHATE ISOMERASE
1R2S 2003-09-29T00:00:00+0000 2.85 CRYSTAL STRUCTURE OF RABBIT MUSCLE TRIOSEPHOSPHATE ISOMERASE
1R2T 2003-09-29T00:00:00+0000 2.25 CRYSTAL STRUCTURE OF RABBIT MUSCLE TRIOSEPHOSPHATE ISOMERASE
4OWG 2014-02-01T00:00:00+0000 1.55 Crystal structure of rabbit muscle triosephosphate isomerase-PEP complex
1HTI 1994-10-12T00:00:00+0000 2.8 CRYSTAL STRUCTURE OF RECOMBINANT HUMAN TRIOSEPHOSPHATE ISOMERASE AT 2.8 ANGSTROMS RESOLUTION. TRIOSEPHOSPHATE ISOMERASE RELATED HUMAN GENETIC DISORDERS AND COMPARISON WITH THE TRYPANOSOMAL ENZYME
1KLG 2001-12-11T00:00:00+0000 2.4 Crystal structure of HLA-DR1/TPI(23-37, Thr28-->Ile mutant) complexed with staphylococcal enterotoxin C3 variant 3B2 (SEC3-3B2)
1KLU 2001-12-12T00:00:00+0000 1.93 Crystal structure of HLA-DR1/TPI(23-37) complexed with staphylococcal enterotoxin C3 variant 3B2 (SEC3-3B2)
1WYI 2005-02-14T00:00:00+0000 2.2 human triosephosphate isomerase of new crystal form
2IAM 2006-09-08T00:00:00+0000 2.8 Structural basis for recognition of mutant self by a tumor-specific, MHC class II-restricted TCR
2IAN 2006-09-08T00:00:00+0000 2.8 Structural basis for recognition of mutant self by a tumor-specific, MHC class II-restricted TCR

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
93.5 Triosephosphate isomerase Q29371 TPIS_PIG
95.2 Triosephosphate isomerase P48500 TPIS_RAT
96.0 Triosephosphate isomerase P17751 TPIS_MOUSE
97.6 Triosephosphate isomerase Q5E956 TPIS_BOVIN
98.0 Triosephosphate isomerase Q5R928 TPIS_PONAB
97.6 Triosephosphate isomerase Q2QD07 TPIS_GORGO
98.4 Triosephosphate isomerase P60175 TPIS_PANTR
98.4 Triosephosphate isomerase P60174 TPIS_HUMAN
98.4 Triosephosphate isomerase P15426 TPIS_MACMU
98.4 Triosephosphate isomerase Q60HC9 TPIS_MACFA
99.2 Triosephosphate isomerase P54714 TPIS_CANLF
100.0 Triosephosphate isomerase P00939 TPIS_RABIT