A structural and dynamic investigation of the inhibition of catalase by nitric oxide

DOI: 10.1039/c3ob41977k
PMID: 24121528

Authors: Marco Candelaresi (University of Strathclyde) , Andrea Gumiero (Diamond Light Source) , Katrin Adamczyk (University of Strathclyde) , Kirsty Robb (Diamond Light Source) , César Bellota-Antón (University of Strathclyde) , Vartul Sangal (University of Strathclyde) , John Munnoch (University of East Anglia) , Gregory M. Greetham (Central Laser Facility, Research Complex at Harwell) , Mike Towrie (Central Laser Facility, Research Complex at Harwell) , Paul A. Hoskisson (University of Strathclyde) , Anthony W. Parker (Central Laser Facility, Research Complex at Harwell) , Nicholas P. Tucker (University of Strathclyde) , Martin A. Walsh (Diamond Light Source) , Neil T. Hunt (University of Strathclyde)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Organic & Biomolecular Chemistry , VOL 11 (44)

State: Published (Approved)
Published: October 2013

Abstract: Determining the chemical and structural modifications occurring within a protein during fundamental processes such as ligand or substrate binding is essential to building up a complete picture of biological function. Currently, significant unanswered questions relate to the way in which protein structural dynamics fit within the structure–function relationship and to the functional role, if any, of bound water molecules in the active site. Addressing these questions requires a multidisciplinary approach and complementary experimental techniques that, in combination, enhance our understanding of the complexities of protein chemistry. We exemplify this philosophy by applying both physical and biological approaches to investigate the active site chemistry that contributes to the inhibition of the Corynebacterium glutamicum catalase enzyme by nitric oxide. Ultrafast two-dimensional infrared spectroscopy (2D-IR) experiments exploit the NO ligand as a local probe of the active site molecular environment and shows that catalase displays a dynamically-restricted, ‘tight,’ structure. X-ray crystallography studies of C. glutamicum catalase confirm the presence of a conserved chain of hydrogen-bonded bound water molecules that link the NO ligand and the protein scaffold. This combination of bound water and restricted dynamics stands in stark contrast to other haem proteins, such as myoglobin, that exhibit ligand transport functionality despite the presence of a similar distal architecture in close proximity to the ligand. We conclude not only that the bound water molecules in the catalase active site play an important role in molecular recognition of NO but also may be part of the mechanistic operation of this important enzyme.

Journal Keywords: Crystallography; X-Ray; Models; Molecular; Nitric; Protein; Spectrophotometry; Infrared; Spectroscopy; Fourier Transform Infrared

Subject Areas: Biology and Bio-materials, Chemistry, Medicine


Instruments: I04-1-Macromolecular Crystallography (fixed wavelength) , I04-Macromolecular Crystallography , I24-Microfocus Macromolecular Crystallography

Added On: 12/11/2013 13:11

Discipline Tags:

Health & Wellbeing Biochemistry Chemistry Structural biology Drug Discovery Life Sciences & Biotech

Technical Tags:

Diffraction Macromolecular Crystallography (MX)