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.
Submitter: Shu-Ching Ou
Submission Date: Jan. 17, 2019, 3:27 p.m.
C41->C43, A62->A64, G72->G74, E104->E106, G122->G124, V154->V156, I170->I172, V231->V233, F240->F242
|Number of data points||97|
|Assays/Quantities/Protocols||Experimental Assay: Dimeric stability ; Experimental Assay: Tm ; Experimental Assay: Km ; Experimental Assay: kcat ; Experimental Assay: Yield ; Derived Quantity: SD of Dimeric stability ; Derived Quantity: SD of Tm ; Derived Quantity: Relative kcat/Km ; Derived Quantity: kcat/Km ; Derived Quantity: SD of Km|
|Libraries||Variants for HsTIM ; Variants for HsTIM_GAP|
|Percent Identity||Matching Chains||Protein||Accession||Entry Name|