Differential effects on enzyme stability and kinetic parameters of mutants related to human triosephosphate isomerase deficiency.


Abstract

Human triosephosphate isomerase (TIM) deficiency is a very rare disease, but there are several mutations reported to be causing the illness. In this work, we produced nine recombinant human triosephosphate isomerases which have the mutations reported to produce TIM deficiency. These enzymes were characterized biophysically and biochemically to determine their kinetic and stability parameters, and also to substitute TIM activity in supporting the growth of an Escherichia coli strain lacking the tim gene. Our results allowed us to rate the deleteriousness of the human TIM mutants based on the type and severity of the alterations observed, to classify four "unknown severity mutants" with altered residues in positions 62, 72, 122 and 154 and to explain in structural terms the mutation V231M, the most affected mutant from the kinetic point of view and the only homozygous mutation reported besides E104D.

Submission Details

ID: uiAUwBD64

Submitter: Shu-Ching Ou

Submission Date: Jan. 17, 2019, 3:27 p.m.

Version: 1

Publication Details
Cabrera N;Torres-Larios A;García-Torres I;Enríquez-Flores S;Perez-Montfort R,Biochim Biophys Acta Gen Subj (2018) Differential effects on enzyme stability and kinetic parameters of mutants related to human triosephosphate isomerase deficiency. PMID:29571745
Additional Information

C41->C43, A62->A64, G72->G74, E104->E106, G122->G124, V154->V156, I170->I172, V231->V233, F240->F242

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
1R2T 2003-09-29T00:00:00+0000 2.25 CRYSTAL STRUCTURE OF RABBIT MUSCLE TRIOSEPHOSPHATE ISOMERASE
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
4OWG 2014-02-01T00:00:00+0000 1.55 Crystal structure of rabbit muscle triosephosphate isomerase-PEP complex
1TIM 1976-09-01T00:00:00+0000 2.5 STRUCTURE OF TRIOSE PHOSPHATE ISOMERASE FROM CHICKEN MUSCLE
8TIM 1998-08-25T00:00:00+0000 2.5 TRIOSE PHOSPHATE ISOMERASE
1TPV 1994-11-07T00:00:00+0000 1.9 S96P CHANGE IS A SECOND-SITE SUPPRESSOR FOR H95N SLUGGISH MUTANT TRIOSEPHOSPHATE ISOMERASE
1TPW 1994-11-07T00:00:00+0000 1.9 TRIOSEPHOSPHATE ISOMERASE DRINKS WATER TO KEEP HEALTHY
1TPU 1994-11-07T00:00:00+0000 1.9 S96P CHANGE IS A SECOND-SITE SUPPRESSOR FOR H95N SLUGGISH MUTANT TRIOSEPHOSPHATE ISOMERASE
1TPH 1993-12-22T00:00:00+0000 1.8 1.8 ANGSTROMS CRYSTAL STRUCTURE OF WILD TYPE CHICKEN TRIOSEPHOSPHATE ISOMERASE-PHOSPHOGLYCOLOHYDROXAMATE COMPLEX

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
180.4 A,B Triosephosphate isomerase P00940 TPIS_CHICK
187.0 A,B Triosephosphate isomerase Q29371 TPIS_PIG
188.8 A,B Triosephosphate isomerase P48500 TPIS_RAT
192.0 A,B Triosephosphate isomerase P17751 TPIS_MOUSE
195.2 A,B Triosephosphate isomerase Q5E956 TPIS_BOVIN
196.8 A,B Triosephosphate isomerase P54714 TPIS_CANLF
196.8 A,B Triosephosphate isomerase P00939 TPIS_RABIT
199.2 A,B Triosephosphate isomerase Q5R928 TPIS_PONAB
198.4 A,B Triosephosphate isomerase Q2QD07 TPIS_GORGO
198.4 A,B Triosephosphate isomerase P15426 TPIS_MACMU
198.4 A,B Triosephosphate isomerase Q60HC9 TPIS_MACFA
200.0 A,B Triosephosphate isomerase P60175 TPIS_PANTR
200.0 A,B Triosephosphate isomerase P60174 TPIS_HUMAN