A National Cancer Institute-designated Comprehensive Cancer Center

Make an appointment: 800-826-HOPE

X-ray Crystallography Core Facility

X-ray Crystallography Core Facility
The X-ray Crystallography Core Facility is one of the Shared Resources facilities at Beckman Research Institute of City of Hope.
 
The objective of the facility is to:
 
  1. Provide structural information to understand mechanisms of biological systems at the atomic level;
  2. Validate binding sites of lead compounds of therapeutic interest;
  3. Facilitate lead discovery through co-crystallization of therapeutic targets and small molecule libraries.
     
Services provided include:
 
  • Sample analysis
  • Crystallization
  • Diffraction quality
  • Data collections
  • Structure determination
 
Contact Us:
 
John C. Williams, Ph.D., Director
Phone:  626-256-HOPE (4673), ext. 60227
City of Hope
1500 East Duarte Road
Duarte, CA  91010-3000
Flower Building, Room 148
 
Research reported in this publication included work performed in the X-ray Crystallography Core supported by the National Cancer Institute of the National Institutes of Health under award number P30CA33572. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.

Services

The X-ray crystallography Core provides a state-of-the-art facility for the generation of crystals and structure determination of macromolecules including proteins, DNA, RNA, and complexes between macromolecules and their ligands.
 
Sample Analysis:
 
Consultation andin silicoanalysis
  • Protein expression and purification
  • Domain structure, including disorder and secondary structure prediction
 
Physical Analysis
 
  • Native PAGE using GE HealthSciences PhastGel
  • Limited Proteolysis and characterization by SDS-PAGE
  • Size Exclusion Chromatography
  • Sedimentation Equilibrium analysis by AUC
  • Sedimentation Velocity analysis by AUC.
 
Crystallization:
 
  1. Set up crystallization trials at 4 ºC and/or 20 ºC.
    a.Initial trials (e.g., appropriate concentrations)
    b.Full scale trials (4 different 96 well factorialsat 3 protein concentrations and 2 temperatures)
     
  2. Optimization – additive screens and factorial overlays
     
  3. Automated Screening at 2 temperatures (scan daily forfirst week, weekly thereafter, finish experiment at 3 months)
 
Diffraction quality:
 
  1. Test diffraction using capillary mounted crystal
     
  2. Test/screen cryo-conditions
 
Data collection:
 
  1. Collect, reduce and merge data. Generate table of statistics
  2. MAD/SAD phasing – help design, collect, reduce and merge MAD data
  3. Generate table of statistics including anomalous dispersion differences
 
Structure Determination:
 
  1. Solve structure by Molecular Replacement
  2. Solve structure by MAD/SAD phasing
  3. Refine structure
  4. Produce relevant statistics (e.g., R and Rfree, RMS deviations)
  5. Structural analysis (superpositions, electrostatics, etc)
  6. Deposit Structure at PDB
 

Equipment

The facility houses a Mosquito Crystallization Robot, a Formulatrix imaging robot (with Automated Crystal Screening at 4 ºC and 20 ºC), a Rigaku Micromax 007 with an R-AxisIV++, and an Oxford cryojet and relevant software and computational hardware for structure determination.
 
The facility also houses a Beckman XLI analytical ultracentrifuge and a GE Health Phast Gel system for the characterization of macromolecular properties.
 
Mosquito Crystallization Robot
The mosquito crystallization robot permits hanging drop, sitting drop, and batch methods in a 96 well format using minimal amounts of protein (~16mL/trail). A number of commercial crystallization screens are available (Qiagen, Hampton Research, Jena).
   
Formulatrix Automated Visualization of Crystallization Trials
The core houses two Formulatrix imaging robots for visualization at 4 ºC and 20 ºC.
Users can quickly browse images, score potential hits and follow crystal growth in time.
   
Rigaku X-ray Diffractometer
Micromax 007 HF, R-axis V++, and Oxford cryojets allow full structure determination.
   
Beckman XL-I Proteomelab
The Beckman Analytical ultracentrifuge affords accurate measurement of hydrodynamic properties, including the association constant of macromolecular complexes, as well as small molecule-macromolecular interactions
 
 

Abstract for Grants

The X-ray core atBeckman Research Institute of City of Hope is a state-of-the-art crystallization and X-ray facility. This facility houses a Mosquito Crystallization robot that uses 50 to 100 nL volumes and permits three different crystallization formats; hanging drop, sitting drops or batch methods under oil. The facility has multiple commercial crystallization factorials (e.g., Hampton, Jena, Nextal), that are frequently used, such that they are always fresh. It has also developed protocols for rapid optimization of individual conditions.
 
To follow crystallization trials, two Formulatrix Imaging Robots are used that automatically image each drop, according to a preset schedule (e.g., nightly). This allows follow-up of crystallization trends at two temperatures and produces a visual record for analysis.
 
For diffraction studies, a Rigaku 007HF generator with an R-axis IV++ imager has been purchased. An Oxford cryojet permits data collection at 100 K to reduce radiation damage.
 
The facility has all the software and computational hardware necessary to determine structures of macromolecular complexes.
 

Pricing

Prices and availability vary. Please contact us for current information.
 
If you are a City of Hope employee, please visit the new intranet site for pricing.

 

 
Research Shared Services

City of Hope embodies the spirit of scientific collaboration by sharing services and core facilities with colleagues here and around the world.
 

Recognized nationwide for its innovative biomedical research, City of Hope's Beckman Research Institute is home to some of the most tenacious and creative minds in science.
City of Hope is one of only 41 Comprehensive Cancer Centers in the country, the highest designation awarded by the National Cancer Institute to institutions that lead the way in cancer research, treatment, prevention and professional education.
Learn more about City of Hope's institutional distinctions, breakthrough innovations and collaborations.
Support Our Research
By giving to City of Hope, you support breakthrough discoveries in laboratory research that translate into lifesaving treatments for patients with cancer and other serious diseases.
 
 
 
 
Media Inquiries/Social Media

For media inquiries contact:

Dominique Grignetti
800-888-5323
dgrignetti@coh.org

 

For sponsorships inquiries please contact:

Stefanie Sprester
213-241-7160
ssprester@coh.org

Christine Nassr
213-241-7112
cnassr@coh.org

 
CONNECT WITH US
Facebook  Twitter  YouTube  Blog
 
NEWS & UPDATES
  • For breast cancer survivors, a common worry is a recurrence of their cancer. Currently, these patients are screened with regular mammograms, but there’s no way to tell who is more likely to have a recurrence and who is fully cleared of her cancer. A new blood test – reported in Cancer Research, a journal of the...
  • Metastasis — the spreading of cancer cells from a primary tumor site to other parts of the body — generally leads to poorer outcomes for patients, so oncologists and researchers are constantly seeking new ways to detect and thwart this malicious process. Now City of Hope researchers may have identified a substa...
  • Deodorant, plastic bottles, grilled foods, artificial sweeteners, soy products … Do any of these products really cause cancer? With so many cancer myths and urban legends out there, why not ask the experts? They can debunk cancer myths while sharing cancer facts that matter, such as risk factors, preventi...
  • Cancer risk varies by ethnicity, as does the risk of cancer-related death. But the size of those differences can be surprising, highlighting the health disparities that exist among various ethnic groups in the United States. Both cancer incidence and death rates for men are highest among African-Americans, acco...
  • George Winston, known worldwide for his impressionistic, genre-defying music, considers music to be his first language, and admits he often stumbles over words – especially when he attempts languages other than English. There’s one German phrase he’s determined to perfect, however: danke schön. Winston thinks h...
  • Few decisions are more important than those involving health care, and few decisions can have such lasting impact, not only on oneself but on relatives and loved ones. Those choices, especially, should be made in advance – carefully, deliberately, free of pain and stress, and with much weighing of values and pr...
  • Using a card game to make decisions about health care, especially as those decisions relate to the end of life, would seem to be a poor idea. It isn’t. The GoWish Game makes those overwhelming, but all-important decisions not just easy, but natural. On each card of the 36-card deck is listed what seriously ill,...
  • Young adults and adolescents with cancer face unique challenges both during their treatment and afterward. Not only are therapies for children and older adults not always appropriate for them, they also must come to terms with the disease and treatment’s impact on their relationships, finances, school or ...
  • Breast cancer is the most common cancer, other than skin cancer, among women in the United States. It’s also the second-leading cause of cancer death, behind lung cancer. In the past several years, various task force recommendations and studies have questioned the benefits of broad screening guidelines fo...
  • Paternal age and the health effects it has on potential offspring have been the focus of many studies, but few have examined the effect parental age has on the risk of adult-onset hormone-related cancers (breast cancer, ovarian cancer and endometrial cancer). A team of City of Hope researchers, lead by Yani Lu,...
  • Hormone therapy, which is prescribed to women for relief of menopausal symptoms such hot flashes, night sweats and vaginal dryness, has recently seen a decline in popularity (and use) due to its link to an increased risk of breast and endometrial cancer. But City of Hope researchers have found that menopausal h...
  • Myeloproliferative neoplasms can’t be narrowed down to a single cancer, but they can be described by a defining characteristic: too many blood cells. The diseases bring with them a host of frustrating, potentially life-altering symptoms, and management of the diseases and their symptoms is crucial. An upcoming ...
  • More than 18,000 researchers, clinicians, advocates and other professionals will convene at the 105th American Association for Cancer Research (AACR) annual meeting taking place in San Diego from April 5 to 9. With more than 6,000 findings being presented over this five-day period, the amount of information can...
  • Cancer of the prostate is the No. 2 cancer killer of men, behind lung cancer, accounting for more than 29,000 deaths annually in this country. But because prostate cancer advances slowly, good prostate health and early detection can make all the difference. Many prostate cancer tumors don’t require immedi...
  • Despite advances made in detecting and treating nonsmall cell lung cancer, its prognosis remains grim. Even patients whose cancers are caught at their earliest stage have only a 50 percent chance of five-year survival. This poor prognosis is due in part to the cancer’s ability to resist treatment, renderi...