Rovnyak Lab NMR Undergraduate Laboratory Page

Uniform ¹⁵N-enriched human ubiquitin : Human Ubiquitin at VLI-Research

The ultimate online resource for NMR data on human ubiquitin is Dr. Richard Harris' Ubiquitin NMR Resource Page

Links around Bucknell
My web page
Bucknell Chemistry Department
Bucknell University

This page contains three modules for NMR in education that we have developed. The first is an example of exploiting some benefits of lower fields and is:

1. ethanol/water exchange experiment on a EFT-60

Two are focused on isotopically enriched proteins, which are commercially available and we show how to incorporate modern biomolecular NMR experiments into undergraduate curricula.

Specifically, we developed laboratories that use NMR instruments configured for typical chemistry applications such as the Bruker ARX 300 MHz Spectrometer with a X{H} probe :

2. Intro to Biomolecular NMR: ¹⁵N Direct Detected HSQC
3. Protein Hydrodynamics: ¹⁵N Relaxation (T₁ and T₂)

1. Ethanol/Water on an EFT-60

An ethanol:water experiment to measure the proton lifetimes on a 60 MHz pulsed FT-NMR (Anasazi Inc.). Here is the application note (copyright G. Manley, D. Rovnyak, 2006).

Ethanol/Water Application Note

2. ¹⁵N Direct Detected HSQC

Supporting material for the implementation of a ¹⁵N-detected ¹H-¹⁵N correlation experiment (HSQC - heteronuclear single quantum coherence transfer) in Chemistry 340, "Biophysical Chemistry", which is described in

Rovnyak, D., Thompson, L.E.,."An accessible two-dimensional solution nuclear magnetic resonance experiment on human ubiquitin", Biochem. Mol. Biol. Ed. 33 (2), 117-122 (2005).

This experiment includes the following material:

a) A set of exercises for discussion or assignment.* Download
b) Raw data and nmrPipe processing scripts for all data.# Download
c) Pulse program (Bruker ARX 300). Download

* A copy with answers is gladly provided to instructors upon request - phone or email (drovnyak@bucknell.edu).
# Tarred an zipped for unix (e.g. linux). First unzip then do 'tar -xvf'.

3. ¹⁵N Direct Detected relaxation (T₁ and T₂) for hydrodynamics

Supporting material for the implementation of ¹⁵N detected relaxation experiments on human ubiquitin for determination of rotational correlation time and protein diameter in Chemistry 340,"Biophysical Chemistry", which is described in a paper to be submitted to Biochemistry and Molecular Biology Education:

Thompson, L.E., Rovnyak, D.,"Accessible NMR Experiments Studying the Hydrodynamics of ¹⁵N-enriched Ubiquitin at Low Fields", submitted (2006).

This experiment includes the following material:

a) T₁ pulse sequence (Bruker Xwinnmr 2.6).* Download
b) T₂ (CPMG) pulse sequence (Bruker Xwinnmr 2.6). * Download
c) MS Excel file to fit T₁/T₂ data to a rotational correlation time. # Download
d) MS Word file: "NMR in 8 Easy Steps!". Download

* Pulse sequence files contain many notes on configuration and use.
# Instructions are built into the excel file.

David Rovnyak Assistant Professor
Department of Chemistry
Bucknell University
Lewisburg, PA 17837
Office : 570-577-3676
FAX : 570-577-1739 (chem dept.)