Abstract
In homodimeric triosephosphate isomerase from Trypanosoma brucei (TbTIM), cysteine 14 of each the two subunits forms part of the dimer interface. This residue is central for the catalysis and stability of TbTIM. Cys14 was changed to the other 19 amino acids to determine the characteristics that the residue must have to yield catalytically competent stable enzymes. C14A, C14S, C14P, C14T, and C14V TbTIMs were essentially wild type in activity and stability. Mutants with Asn, Arg, and Gly had low activities and stabilities. The other mutants had less than 1% of the activity of TbTIM. One of the latter enzymes (C14F) was purified to homogeneity. Size exclusion chromatography and equilibrium sedimentation studies showed that C14F TbTIM is a monomer, with a k(cat) approximately 1000 times lower and a K(m) approximately 6 times higher than those of TbTIM. In C14F TbTIM, the ratio of the elimination (methylglyoxal and phosphate formation) to isomerization reactions was higher than in TbTIM. Its secondary structure was very similar to that of TbTIM; however, the quantum yield of its aromatic residues was lower. The analysis of the data with the 19 mutants showed that to yield enzymes similar to the wild type, the residue must have low ...Continue Reading
References
Aug 20, 1991·Journal of Molecular Biology·R K WierengaW G Hol
Sep 1, 1973·The Biochemical Journal·S G Waley
Dec 1, 1981·Biophysical Journal·M L JohnsonH R Halvorson
Jul 3, 1995·FEBS Letters·T V BorchertR K Wierenga
Feb 1, 1994·Experimental Parasitology·M R MowattT E Nash
Feb 1, 1993·European Journal of Biochemistry·T V BorchertR K Wierenga
Mar 29, 1996·Journal of Molecular Biology·V MainfroidK Goraj
Oct 1, 1996·European Journal of Biochemistry·G Garza-RamosA Gómez-Puyou
Jan 1, 1997·Biophysical Journal·J S Philo
Jul 1, 1997·European Journal of Biochemistry·A LandaD A Fernández-Velasco
Jan 27, 1998·The Journal of Biological Chemistry·M AlvarezD Maes
Jul 8, 1998·European Journal of Biochemistry·G Garza-RamosA Gómez-Puyou
Oct 8, 1998·Journal of Molecular Biology·E MaldonadoR Perez-Montfort
Mar 13, 2001·Journal of Molecular Biology·H WaldenG L Taylor
Apr 5, 2001·Journal of Molecular Biology·J XiangN S Sampson
Jul 18, 2001·FEBS Letters·S K SinghP Balaram
Nov 15, 1993·Structure·T V BorchertR K Wierenga
Citations
Jul 20, 2002·Biochemical and Biophysical Research Communications·Alfredo Téllez-ValenciaArmando Gómez-Puyou
Feb 4, 2011·Protein Engineering, Design & Selection : PEDS·Mousumi BanerjeeP Balaram
Mar 15, 2011·Insect Biochemistry and Molecular Biology·Jorge MoraesCarlos Logullo
Jun 22, 2010·Molecular and Cellular Probes·Matthew J NolanRobin B Gasser
Dec 14, 2007·Molecular and Biochemical Parasitology·Sergio Enriquez-FloresHoracio Reyes-Vivas
May 18, 2004·Proteins·Gabriel López-VelázquezHoracio Reyes-Vivas
Jan 16, 2007·Proteins·Viviana Zomosa-SignoretArmando Gómez-Puyou
Jun 5, 2013·Proteins·Samuel Lara-GonzálezClaudia G Benítez-Cardoza
Nov 11, 2006·Journal of Molecular Biology·Horacio Reyes-VivasGabriel Lopez-Velazquez
Sep 19, 2006·FEMS Microbiology Letters·Divya MathurLalit C Garg
Apr 6, 2017·Proteins·Vanesa Olivares-IllanaArmando Gómez-Puyou
Oct 25, 2007·Proteins·Chen-Hsi ChuYuh-Ju Sun
Dec 4, 2012·International Journal of Molecular Sciences·Luiz SaramagoJorge Moraes
Dec 22, 2016·Frontiers in Plant Science·Laura M López-CastilloLuis G Brieba
Oct 1, 2018·Biochemical and Biophysical Research Communications·Timothy C ChangJason P Schwans
Oct 8, 2008·Biochemistry·Luis A TellezD Alejandro Fernández-Velasco
Feb 27, 2008·Biochemistry·Nallely CabreraRuy Perez-Montfort
Sep 24, 2021·Science·Tao CaiYanhe Ma