Dynamics and hydration explain failed functional transformation in dehalogenase design.

نویسندگان

  • Jan Sykora
  • Jan Brezovsky
  • Tana Koudelakova
  • Maryna Lahoda
  • Andrea Fortova
  • Tatsiana Chernovets
  • Radka Chaloupkova
  • Veronika Stepankova
  • Zbynek Prokop
  • Ivana Kuta Smatanova
  • Martin Hof
  • Jiri Damborsky
چکیده

We emphasize the importance of dynamics and hydration for enzymatic catalysis and protein design by transplanting the active site from a haloalkane dehalogenase with high enantioselectivity to nonselective dehalogenase. Protein crystallography confirms that the active site geometry of the redesigned dehalogenase matches that of the target, but its enantioselectivity remains low. Time-dependent fluorescence shifts and computer simulations revealed that dynamics and hydration at the tunnel mouth differ substantially between the redesigned and target dehalogenase.

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

منابع مشابه

Interchange of catalytic activity within the 2-enoyl-coenzyme A hydratase/isomerase superfamily based on a common active site template.

The structures and chemical pathways associated with the members of the 2-enoyl-CoA hydratase/isomerase enzyme superfamily are compared to show that a common active site design provides the members of this family with a CoA binding site, an expandable acyl binding pocket, an oxyanion hole for binding/polarizing the thioester C=O, and multiple active site stations for the positioning of acidic a...

متن کامل

Ruminal Kinetics of Nutrients Degradation, Hydration, and Functional Specific Gravity of Three Types of Beet Pulp

Two experiments were conducted to evaluate the relationships between nutrients degradability, kinetics of hydration, functional specific gravity (FSG) of the three types of beet pulp (BP) including fine (FBP), normal (NBP) and pelleted (PBP) BP. In experiment 1, about 3 g of samples was weighed in sealed nylon bags (6 cm×7.5 cm, 40±5 µm pore size), incubated in rumen of two cannulated Holstein ...

متن کامل

Propensity based classification: Dehalogenase and non-dehalogenase enzymes

The present work was designed to classify and differentiate between the dehalogenase enzyme to non–dehalogenases (other hydrolases) by taking the amino acid propensity at the core, surface and both the parts. The data sets were made on an individual basis by selecting the 3D structures of protein available in the PDB (Protein Data Bank). The prediction of the core amino acid were predicted by I...

متن کامل

Molecular Docking Based on Virtual Screening, Molecular Dynamics and Atoms in Molecules Studies to Identify the Potential Human Epidermal Receptor 2 Intracellular Domain Inhibitors

Human epidermal growth factor receptor 2 (HER2) is a member of the epidermal growth factor receptor family having tyrosine kinase activity. Overexpression of HER2 usually causes malignant transformation of cells and is responsible for the breast cancer. In this work, the virtual screening, molecular docking, quantum mechanics and molecular dynamics methods were employed to study protein–ligand ...

متن کامل

Estimation of the Elastic Properties of Important Calcium Silicate Hydrates in Nano Scale - a Molecular Dynamics Approach

Approximately, 50 to 70 percent of hydration products in hydrated cement paste are polymorphisms of C-S-H gel. It is highly influential in the final properties of hardened cement paste. Distinguishing C-S-H nano-structure significantly leads to determine its macro scale ensemble properties. This paper is dealt with nano-scale modeling. To achieve this, the most important C-S-H compounds, with a...

متن کامل

ذخیره در منابع من


  با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

عنوان ژورنال:
  • Nature chemical biology

دوره 10 6  شماره 

صفحات  -

تاریخ انتشار 2014