Nuclear Magnetic Resonance NMR-based Structural Biology Research

Our laboratory specializes in nuclear magnetic resonance (NMR)-based structural biology research. We are particularly interested in understanding the crucial links between the molecular structures, internal dynamics, and biochemical functions of proteins that are of importance to human endeavors. Questions of interest are: What is the connection between a protein’s three-dimensional architecture, flexibility of its amino acids and of its structural elements, and its biological function(s)? How do atomic structures and internal dynamics modulate the biochemical activity of proteins? What is the significance of conserved amino acid residues in protein families? Our approach to providing answers to these scientific issues is to use modern multidimensional (2D, 3D, 4D), heteronuclear (¹H, ¹⁵N, ¹³C, ²H) solution nuclear magnetic resonance (NMR) spectroscopy in conjunction with complementary biophysical techniques. We are currently investigating the structural and functional properties of several intriguing proteins.

Our Thermal Biology-related research is focused on the characterization of extremophilic proteins from microorganisms existing in the thermal environments in Yellowstone National Park. A research theme that is particularly attractive to us is: What are the characteristic molecular features of proteins originating from thermophilic organisms that distinguish them from their mesophilic partners? For example, how do organisms adapt to the high temperature (T > 70^oC), acidic (pH < 4.0), and “toxic” metal-rich (arsenic, iron, copper, mercury, and others) environment of YNP thermal pools? How do proteins originating from extremophiles get modified to function in such an environmental context? What changes take place at the molecular levels? How do proteins “modify” their thermodynamical characteristics (i.e. thermal stability, flexibility of functional residues) to operate efficiently in thermophilic conditions? How do proteins “cope” with arsenic, cadmium, or copper-rich environments? Modern multidimensional, heteronuclear (¹H, ¹⁵N, ¹³C, ²H) solution nuclear magnetic resonance (NMR) spectroscopy is an excellent technique to provide answers to these fundamental issues, both at the structural and motional levels of atoms, and complements well the X-ray-based structural genomics research taking place within TBI.

Selected publications:

Internal dynamics of the tryptophan repressor (TrpR) and two functionally distinct TrpR variants, L75F-TrpR and A77V-TrpR, in their l-Trp-bound forms.
Tripet BP, Goel A, Copie V. Biochemistry. 2011 Jun 14;50(23):5140-53. Epub 2011 May 20.

Structural and functional characterization of an archaeal clustered regularly interspaced short palindromic repeat (CRISPR)-associated complex for antiviral defense (CASCADE).
Lintner NG, Kerou M, Brumfield SK, Graham S, Liu H, Naismith JH, Sdano M, Peng N, She Q, Copié V, Young MJ, White MF, Lawrence CM. J Biol Chem. 2011 Jun 17;286(24):21643-56. Epub 2011 Apr 20.

The structure of the CRISPR-associated protein Csa3 provides insight into the regulation of the CRISPR/Cas system.
Lintner NG, Frankel KA, Tsutakawa SE, Alsbury DL, Copié V, Young MJ, Tainer JA, Lawrence CM. J Mol Biol. 2011 Jan 28;405(4):939-55. Epub 2010 Nov 18.

Backbone amide dynamics studies of Apo-L75F-TrpR, a temperature-sensitive mutant of the tryptophan repressor protein (TrpR): comparison with the (15)N NMR relaxation profiles of wild-type and A77V mutant Apo-TrpR repressors.
Goel A, Tripet BP, Tyler RC, Nebert LD, Copié V.
Biochemistry. 2010 Sep 21;49(37):8006-19.

(1)H, (13)C, (15)N backbone and side chain NMR resonance assignments for E73 from Sulfolobus spindle-shaped virus ragged hills, a hyperthermophilic crenarchaeal virus from Yellowstone National Park.
Schlenker C, Menon S, Lawrence CM, Copié V.
Biomol NMR Assign. 2009 Dec;3(2):219-22. Epub 2009 Jul 16.

Current Laboratory Personnel:

Anupam Goel: Ph.D. Student
Emilio Reyes, Ph.D. Student
Casey Schlenker, Ph.D. Student
Haley Arthun, Undergraduate Student
Woody Cranston, Technical Support Staff

Return to regular view	Text-only