LYSOZYME STRUCTURE

Chem 112L

Biophysical and Bioanalytical Laboratory

LYSOZYME CRYSTAL

Lecturer

Dr. Kalju Kahn
Office: PSB-N 2623
Office hours: Tue 3:30-4:30 PM, "Open Door" policy other times
Phone: 893-6157
E-mail: kalju@chem.ucsb.edu
Website: http://www.chem.ucsb.edu/~kalju

Teaching Assistants

MW2:		Andrew Benkovitch (abenkovichr@chem.ucsb.edu)
		Office hours: Chem     Phone: x 
TR6:	 	Paul Zakrevsky (pzakrevsky@chem.ucsb.edu) 
		Office hours: Chem     Phone: x 

Mission statement

Chem 112L is an undergraduate laboratory course where students use advanced instrumental techniques to study the structure and function of biological molecules. Students will work on seven projects and learn to use techniques such as circular dichroism, nuclear magnetic resonance spectroscopy, biological mass spectrometry, differential UV spectrophotometry, protein crystallography, and computational biochemistry.

Syllabus and Schedules

       
      Lab section 1: Mon 2:00-5:50; Wed 2:00-5:50;
      Lab section 2: Tue 6:00-9:50; Thu 6:00-9:50; 
        
      Lab sections are in PSB-N 2619 or Chem 1153 (SGI lab)   
Syllabus General information about the course; course schedule PDF
History Course website for Chem 112L (Spring 2009) Link
Grading Grading Rubrics DOC
Upload Submit your assignments as PDF files Link
Exams How to prepare for the first midterm PDF
Exams Selected CHEM 112C Lecture Materials ZIP
Exams How to prepare for the second midterm PDF
Exams Sample Questions for Midterm 1 (2003) PDF
Exams Sample Questions for Midterm 2 (2002) PDF
Exams Midterm 1 Key (Spring 2006) PDF
Exams Midterm 1 Questions PDF
Poster: Poster Presentation Guidelines Link

Laboratory Manual

Students in the class do not have to purchase the laboratory manual. Each chapter of the lab manual can be downloaded here in the PDF format. Please note that you can follow hyperlinks that are in the PDF files by clicking on the link.

Experiments Lab manuals are in the PDF format. Download Adobe Acrobat Here Acrobat
All Theory Manual for Chem 112L PDF
Exp 1: Tutorial on Minimization Link
Exp 1: Conformational analysis of allantoin: Operations Manual PDF
Exp 1: Conformational analysis of allantoin: NMR Instrument Manual PDF
Exp 2 Enzyme Kinetics PDF
Exp 3 Protein Unfolding Monitored by Circular Dichroism PDF
Exp 4 Ligand Binding to Lysozyme PDF
Exp 5 Protein Mass Spectrometry PDF
Math Mathematica Tutorial, Part I from Fall Quarter PDF
Math Mathematica Tutorial, Part II PDF
Math Mathematica Tutorial, Part III PDF
Exp 6 Operation Manual for Protein Crystallography PDF

Supporting Resources and Data

Data files, instrument manuals, and selected lectures notes to support the course are available below

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Supporting Support materials PDF format. Download Adobe Acrobat Here Acrobat
Allantoin Quantum Harmonic Oscillator (using Mathematica) Link
Allantoin Structural Studies of 2',3'-Difluorinated Nucleosides ... PDF
Allantoin Introduction to Monte Carlo Method (UCSB) Link
Allantoin Pathways to a Protein Folding Intermediate... (Duan and Kollman) PDF
Allantoin Molecular Dynamics (Primer by Furio Ercolessi, University of Udine, Italy) Link
Allantoin BOSS User Manual (Jorgensen Research Group at Yale) Link
Allantoin NMR Lecture Notes PDF
Allantoin "Structure Determination of Biological Macromolecules in Solution Using NMR Spectroscopy" by Gerhard Wider PDF
Allantoin "Structure calculation of biological macromolecules from NMR data" by Peter Güntert PDF
Allantoin Ab initio calculation of spin-spin coupling constants Link
Allantoin 1D proton spectrum in Hertz units, sample plot PNG
Allantoin 1D proton spectrum in Hertz units (MW) DATA
Allantoin 1D proton spectrum in Hertz units (TR) DATA
Allantoin Transient NOE spectrum, sample plot PNG
Allantoin Transient NOE spectrum, irradiate at 5.2 ppm DATA
Allantoin Transient NOE spectrum, irradiate at 5.8 ppm DATA
Allantoin Transient NOE spectrum, irradiate at 6.9 ppm DATA
Allantoin Transient NOE spectrum, irradiate at 8.1 ppm DATA
Allantoin Transient NOE spectrum, irradiate at 10.5 ppm DATA
Allantoin COSY (2D) spectrum PNG
Allantoin COSY (2D) spectrum zoom in 5-7 ppm PNG
Allantoin DFQ-COSY (2D) spectrum PNG
Allantoin NOESY (2D) spectrum PNG
Allantoin TOCSY (2D) spectrum PNG
Allantoin Sample NMR Peak Data Files Singlet Doublet
Allantoin Full 1D spectrum collected in water with WATERGATE PNG
Enzymes "Challanges in Enzyme Mechanism and Energetics" PDF
Enzymes "Perspective on "Statistical Analysis of Enzyme Kinetics Data"" PDF
Enzymes "Statistical Analysis of Enzyme Kinetics Data" PDF
Enzymes "Fitting Enzyme-Kinetic Data to V/K" PDF
Enzymes "On the Meaning of Km and V/K " PDF
Enzymes "A Quantitative Approach to Enzyme Inhibition" PDF
Enzymes "Understanding Enzyme Inhibition" by Raymond Ochs PDF
Folding "Protein Stability Curves" by Becktel and Schellman PDF
Folding "Heat Capacity in Proteins" by Prabhu and Sharp PDF
Folding "Synchrotron radiation circular dichroism spectroscopy of proteins ..." PDF
Protein MS Mass Spectrometry Lecture by Dr. Pavlovich PDF
Protein MS "Electrospray: Principles and Practice" by Simon Gaskell PDF
Protein MS "Using Mass Spectrometry for Proteins" by Martha M. Vestling PDF
Protein MS "Principles of FT-ICR and its Application to Structural Biology" by Barrow et. al. PDF
Protein MS: "Proteomic Tools For Quantitation By Mass Spectrometry " by Jennie Lill PDF
MS Mass Spectrometry Data Tables PDF
MS Mass Spectra Figures 1-3 PDF
MS Mass Spectra Figures A-F PDF
Binding "Induction of a Remarkable Conformational Change in a Human Telomeric Sequence ... " PDF
Binding Review of Thermodynamics: Heat Capacity PDF
X-ray Protein Crystallography: Reading Link Link
X-ray Crystallography 101 by Bernhard Rupp at LLNL Link
X-ray "High-resolution neutron protein crystallography ..." PDF
X-ray Electron Density Map for Crystallography (Coot) CCP4
X-ray Electron Density Map for Crystallography (O and Pymol) OMAP
X-ray Amino Acid Residue Positions for Crystallography PDB

Student Files-Protein Crystallography Experiment

One of the experiments in the Biophysical and Bioanalytical Chemistry Laboratory at UCSB is dedicated to protein crystallography and X-ray diffraction. On the first day, students determine the best conditions for growing lyzozyme crystals via hanging drop crystallization trials. Some of the crystals obtained by students are shown below.

MW-A MW-B TR-A TR-B
Crystals from the MW2 Section: Day 1
Crystals from the MW2 Section: Day 2
Crystals from the TR6 Section: Day 1
Crystals from the TR6 Section: Day 2
Crystals from the TR6 Section: Day 3
Crystals from 2009
Crystals from 2008
Crystals from 2007
Crystals from 2006
Crystals from 2005

On the following day, crystals were examined and diffraction data was collected on our in-house X-ray diffractiometer. Many students obtained crystals that were large enough for collecting good-quality diffraction data. Some of our best crystals diffracted down to 2.4 Angstroms, which is an acceptable resolution in modern macromolecular crystallography.

MW-B TR-A TR-B1 TR-B2

On the last day, students analyzed diffraction data to determine the unit cell characteristics and build a partial protein structure into the electron density map. We thank UCSB's Instructional Development Program for funding the development of this experiment; Dr. Alwyn Jones from Uppsala for providing copies of the program "O", and our gradute student Scott Hauenstein from the Perona group for heping implement this experiment.

MW-B TR-A

Physical Biochemistry Links

Electronic Journals
Protein Data Bank
Wolfram Mathematica Features and Tutorials
PyMOL Molecular Visualization Program
Spectroscopy Online Resources
Protein Prospector: Proteomics Mass Spectrometry Database

UCSB links

UCSB General Catalog
UCSB Gold Login
UCSB Umail Access
UCSB Environmental Health and Safety


Course materials by Dr. Kalju Kahn, Department of Chemistry and Biochemistry, UC Santa Barbara. ©2004-2010