• Computer Core
• Crystallization Core
• Protein Expression
• X-ray Crystallography
• NMR Core
• Instrumentation

Instrumentation

The facility provides the UCLA-DOE lab and other members of the UCLA biochemistry community with modern instruments for biomolecule characterization. Instruments are available for biomolecule purification, molecular weight determination, kinetic and thermodynamic analysis of ligand binding, structural characterization, gel documentation and analysis, radioisotope detection and quantification, and oligonucleotide synthesis. The lab manager, Martin Phillips, provides training and assistance in the use of the facility equipment.

Facilities and Equipment

The UCLA-DOE Biochemistry Instrumentation Facility is housed in two adjacent laboratories in Young Hall. The bench tops are wide and deep to easily accommodate large or oddly shaped equipment. Computer network connections are provided throughout the rooms for networking of all the computers controlling the instruments. The room has a curtain to provide darkness for sensitive fluorescence experiments.

Equipment Currently Housed in the Facility:

• Spectrofluorimeter - SPEX: Single photon counting measurements of fluorescence intensity. Equipped with polarizers for fluorescence anisotropy measurements.
• Circular Dichroism Spectrophotometer - JASCO: For the determination of secondary structure content and monitoring structural changes in proteins and nucleic acids. The instrument is equipped with a heating/cooling water bath for monitoring conformational changes as a function of temperature.
• UV/Visible spectrophotometer - Hewlett Packard: This will be used for routine measurements of absorbance spectra and simple concentration measurements. Diode Array detection permits rapid measurement of full spectra. Although this is standard laboratory equipment, it is needed for convenient use of other equipment within the core facility.
• Analytical Ultracentrifuge - Beckman Model XL-A: Currently has absorbance detection only. For sedimentation velocity, sedimentation equilibrium and diffusion coefficient measurements. Provides estimations of molecular weight, shape and binding constants.
• Precision Density Meter - Mettler/Paar DMA O2D: For accurate determination of solution densities and partial specific volume (useful for molecular weight determination).
• *Pharmacia Biacore X: For detecting molecular interactions and determination of equilibrium binding constants and binding and dissociation rate constants.
• *Differential Scanning Calorimeter - Calorimetry Sciences Corp. NANO: For obtaining thermodynamic parameters of thermally induced conformational changes.
• *Titration Calorimeter - Calorimetric Sciences Corporation Jet: For measuring thermodynamic parameters of molecular interactions.
• *Stopped-Flow Reaction Analyzer - Applied Photophysics: For monitoring of fast reactions (instrument dead time <1ms). Possesses both absorbance and fluorescence detection capabilities.
• Protein Purification System - Perseptive Biosystems BioCad 700E: A versatile HPLC system capable of automatically testing a large array of purification methods on all chromatography media. A wide range of flow rates permits use on both analytical and preparative scale columns.
• Phosphorimager - Molecular Dynamics 445 SI: Used for sensitive and quantitative measurement of radioactivity on gels or blots.
• DNA synthesizer - Beckman Oligo 1000M: Oligonucleotide synthesis. Performs 3 simultaneous syntheses with short cycle times.
• Liquid Scintillation Counters - Beckman LS8100 and LS3133P: Radioactivity quantification. (These are very old instruments near the end of their useful life span.)
• *CCD Gel Documentation System: For recording gels and quantification of band intensities.
• *A Wyatt Technology Corp. Three-angle Light Scattering Detector: The instrument will be used to measure molecular weights and therefore oligomerization states of our proteins. It will be used mostly as a chromatography detector, but can also be used in batch mode (direct injection).

Mode of Operation

The laboratory provides biochemists and structural molecular biologists at UCLA with easy access to sophisticated instruments. To access these instruments, they simply need to discuss their needs with the facility Manager, Dr. Phillips, who is responsible for training them in the use of the instrument. In some cases, the facility Manager will simply run the experiment for the investigator. For example, use of the analytical ultracentrifuge and analysis of the results requires significant training. In cases where the investigator only requires the molecular weight determination of a few samples, it is a more efficient use of time for the Manager to run the experiment rather than invest in training. In any case, the best course of action will be decided upon in consultation with the investigator.

At present, instrument time is offered on a first come first served basis to anyone at UCLA. In the event that a particular instrument comes under heavy use, it may become necessary to prioritize access. In this case, priority will be assigned based on financial contributions to the facility in general or the instrument in particular. Members of the DOE laboratory, members of Department of Chemistry and Biochemistry and principle investigators on the shared instrumentation grant for a particular instrument will have priority over all others at this time. If access is severely impacted, purchase of additional instruments of the same kind must be considered. Eventually a recharge system will be set up to recoup the cost of supplies and service contracts.