2.3.1.12 A174S mutation in the inner lipoyl-bearing domain (L2), pyruvate dehydrogenase kinase 2 binding: 69% 672273 2.3.1.12 D164A mutation in the inner lipoyl-bearing domain (L2), pyruvate dehydrogenase kinase 2 binding: 55% 672273 2.3.1.12 D172A mutation in the inner lipoyl-bearing domain (L2), pyruvate dehydrogenase kinase 2 binding: 66% 672273 2.3.1.12 E162A mutation in the inner lipoyl-bearing domain (L2), pyruvate dehydrogenase kinase 2 binding: 93% 672273 2.3.1.12 E179A mutation in the inner lipoyl-bearing domain (L2), pyruvate dehydrogenase kinase 2 binding: 28% 672273 2.3.1.12 E182A binding of mutant L2 domain to pyruvate dehydrogenase phosphatase isozyme 1 is hindered 663011 2.3.1.12 E182A mutation in the inner lipoyl-bearing domain (L2), pyruvate dehydrogenase kinase 2 binding: 100% 672273 2.3.1.12 E182Q binding of mutant L2 domain to pyruvate dehydrogenase phosphatase isozyme 1 is hindered 663011 2.3.1.12 E183A mutation in the inner lipoyl-bearing domain (L2), pyruvate dehydrogenase kinase 2 binding: 98% 672273 2.3.1.12 F231A ability to be post-translationally lipoylated remains 486229 2.3.1.12 G610C 500fold reduced enzymatic activity 486228 2.3.1.12 H602C inactive 736448 2.3.1.12 I176A mutation in the inner lipoyl-bearing domain (L2), pyruvate dehydrogenase kinase 2 binding: not measurable 672273 2.3.1.12 I229A ability to be post-translationally lipoylated remains 486229 2.3.1.12 I229A/F231A ability to be post-translationally lipoylated remains, far-UV CD spectrum differs from wild-type enzyme 486229 2.3.1.12 K136A site-directed mutagenesis, thermodynamics and kinetics in comparison to the wild-type enzyme 661632 2.3.1.12 K153A site-directed mutagenesis, thermodynamics and kinetics in comparison to the wild-type enzyme 661632 2.3.1.12 K173A mutation in the inner lipoyl-bearing domain (L2), pyruvate dehydrogenase kinase 2 binding: not measurable 672273 2.3.1.12 K191E the mutant shows 50% activity compared to the wild type enzyme 725521 2.3.1.12 K191R the mutant shows 79% activity compared to the wild type enzyme 725521 2.3.1.12 K244Q unlipoylatable domain 486166 2.3.1.12 K276A mutant shows negligible binding to human pyruvate dehydrogenase with 86fold higher KD compared to wild-type 686772 2.3.1.12 K96A mutation in potential lipoylation site, mutant protein entirely supports the assignment of Lys96 as the site of lipoylation 720722 2.3.1.12 K96R mutation in potential lipoylation site, mutant protein entirely supports the assignment of Lys96 as the site of lipoylation 720722 2.3.1.12 L140A mutation in the inner lipoyl-bearing domain (L2), pyruvate dehydrogenase kinase 2 binding: not measurable 672273 2.3.1.12 L189A ability to be post-translationally lipoylated remains 486229 2.3.1.12 M131A site-directed mutagenesis, thermodynamics and kinetics in comparison to the wild-type enzyme 661632 2.3.1.12 M194A ability to be post-translationally lipoylated remains 486229 2.3.1.12 M197A ability to be post-translationally lipoylated remains 486229 2.3.1.12 additional information a truncated version of dihydrolipoyl acetyltransferase is created lacking the N-terminal 319 amino acids 690088 2.3.1.12 additional information construction of diverse L2 domain mutants, e.g. substitutions of A172, D173, Leu140, Glu162, Glu181, and Glu179 highly reduce binding of L2 domain to pyruvate dehydrogenase phosphatase isozyme 1 663011 2.3.1.12 additional information deleting the LAT1 gene abolishes life span extension induced by calorie restriction. Overexpressing Lat1 extends life span, and this life span extension is not further increased by calorie restriction. Similar to calorie restriction, life span extension by Lat1 overexpression largely requires mitochondrial respiration. Lat1 overexpression does not require the Sir2 family to extend life span, suggesting that Lat1 mediates a branch of the calorie restriction pathway that functions in parallel to the Sir2 family 674890 2.3.1.12 additional information protein includes a single lipoyl-prosthetic group covalently attached to Lys96. Despite low primary sequence identity with the bacterial enzyme, the plant protein is recognized and modified by Escherichia coli E2 lipoyl-addition system 720722 2.3.1.12 N614D 9fold reduced enzymatic activity 486228 2.3.1.12 R129A the mutant shows 34% activity compared to the wild type enzyme 725521 2.3.1.12 R129E the mutant shows 3.0% activity compared to the wild type enzyme 725521 2.3.1.12 R129K the mutant shows 77% activity compared to the wild type enzyme 725521 2.3.1.12 R130A the mutant shows wild type activity 725521 2.3.1.12 R130E the mutant shows 57% activity compared to the wild type enzyme 725521 2.3.1.12 R135A site-directed mutagenesis, thermodynamics and kinetics in comparison to the wild-type enzyme 661632 2.3.1.12 R135C site-directed mutagenesis, additional alkylation of the mutant enzymes with methyl, ethyl, propyl and butyl groups, the modifications cause alterations in interaction of core unit, E1 and E3 in the enzyme complex compared to the wild-type complex, thermodynamics and kinetics in comparison to the wild-type enzyme, overview 661632 2.3.1.12 R135K site-directed mutagenesis, thermodynamics and kinetics in comparison to the wild-type enzyme 661632 2.3.1.12 R135L site-directed mutagenesis, thermodynamics and kinetics in comparison to the wild-type enzyme 661632 2.3.1.12 R135M site-directed mutagenesis, thermodynamics and kinetics in comparison to the wild-type enzyme 661632 2.3.1.12 R139A site-directed mutagenesis, thermodynamics and kinetics in comparison to the wild-type enzyme 661632 2.3.1.12 R146A site-directed mutagenesis, thermodynamics and kinetics in comparison to the wild-type enzyme 661632 2.3.1.12 R150A the mutant shows 8.0% activity compared to the wild type enzyme 725521 2.3.1.12 R150E the mutant shows 2.4% activity compared to the wild type enzyme 725521 2.3.1.12 R150K the mutant shows 5.0% activity compared to the wild type enzyme 725521 2.3.1.12 R153E the mutant shows 60% activity compared to the wild type enzyme 725521 2.3.1.12 R153K the mutant shows 83% activity compared to the wild type enzyme 725521 2.3.1.12 R156A site-directed mutagenesis, thermodynamics and kinetics in comparison to the wild-type enzyme 661632 2.3.1.12 R161A the mutant shows 65% activity compared to the wild type enzyme 725521 2.3.1.12 R161E the mutant shows 84% activity compared to the wild type enzyme 725521 2.3.1.12 R196A mutation in the inner lipoyl-bearing domain (L2), pyruvate dehydrogenase kinase 2 binding: 79% 672273 2.3.1.12 R202E the mutant shows 10% activity compared to the wild type enzyme 725521 2.3.1.12 R202K the mutant shows 86% activity compared to the wild type enzyme 725521 2.3.1.12 R297A mutant is found to have KD 6.8fold higher than that for the wild-type, indicating a possible involvement of this residue in the interaction with human pyruvate dehydrogenase, but not with the C-terminal residue of beta-subunit 686772 2.3.1.12 S133A site-directed mutagenesis, thermodynamics and kinetics in comparison to the wild-type enzyme 661632 2.3.1.12 S558A 200fold reduced enzymatic activity 486228 2.3.1.12 V180S/E181L binding of mutant L2 domain to pyruvate dehydrogenase phosphatase isozyme 1 is hindered 663011 2.3.1.12 V188A ability to be post-translationally lipoylated remains, far-UV CD spectrum differs from wild-type enzyme 486229