"The Journal of Biological Chemistry" . "A Single Mutation in a Tunnel to the Active Site Changes the Mechanism and Kinetics of Product Release in Haloalkane Dehalogenase LinB"@en . "Many enzymes have buried active sites. The properties of the tunnels connecting the active site with bulk solvent affect ligand binding and unbinding and also the catalytic properties. Here, we investigate ligand passage in the haloalkane dehalogenase enzyme LinB and the effect of replacing leucine by a bulky tryptophan at a tunnel-lining position. Transient kinetic experiments show that the mutation significantly slows down the rate of product release. Moreover, the mechanism of bromide ion release is changed from a one-step process in the wild type enzyme to a two-step process in the mutant. The rate constant of bromide ion release corresponds to the overall steady-state turnover rate constant, suggesting that product release became the rate-limiting step of catalysis in the mutant. We explain the experimental findings by investigating the molecular details of the process computationally."@en . "14310" . "7"^^ . . "000308074600073" . "Many enzymes have buried active sites. The properties of the tunnels connecting the active site with bulk solvent affect ligand binding and unbinding and also the catalytic properties. Here, we investigate ligand passage in the haloalkane dehalogenase enzyme LinB and the effect of replacing leucine by a bulky tryptophan at a tunnel-lining position. Transient kinetic experiments show that the mutation significantly slows down the rate of product release. Moreover, the mechanism of bromide ion release is changed from a one-step process in the wild type enzyme to a two-step process in the mutant. The rate constant of bromide ion release corresponds to the overall steady-state turnover rate constant, suggesting that product release became the rate-limiting step of catalysis in the mutant. We explain the experimental findings by investigating the molecular details of the process computationally." . . . . . "10.1074/jbc.M112.377853" . "Biedermannov\u00E1, Lada" . . . "[1CC491546B0D]" . . "Damborsk\u00FD, Ji\u0159\u00ED" . . "G\u00F3ra, Artur Wiktor" . "4"^^ . "0021-9258" . . . "287" . . "RIV/00216224:14310/12:00057142" . "Kov\u00E1cs, Mih\u00E1ly" . . "HLD; haloalkane dehalogenase; MD; molecular dynamics; RAMD; random acceleration molecular dynamics; ABF; adaptive biasing force; RC; reaction coordinate; FEP; free energy perturbation; NATA; N-acetyltryptophan amide"@en . . "G\u00F3ra, Artur Wiktor" . "RIV/00216224:14310/12:00057142!RIV13-AV0-14310___" . . "A Single Mutation in a Tunnel to the Active Site Changes the Mechanism and Kinetics of Product Release in Haloalkane Dehalogenase LinB" . "US - Spojen\u00E9 st\u00E1ty americk\u00E9" . . "Prokop, Zbyn\u011Bk" . "A Single Mutation in a Tunnel to the Active Site Changes the Mechanism and Kinetics of Product Release in Haloalkane Dehalogenase LinB"@en . "34" . "Wade, Rebecca C." . . . . . "P(ED0001/01/01), P(IAA401630901)" . "120620" . "A Single Mutation in a Tunnel to the Active Site Changes the Mechanism and Kinetics of Product Release in Haloalkane Dehalogenase LinB" . . . "\u0160ebestov\u00E1, Eva" . . . . . . . . . "13"^^ .