The Huck Institutes of the Life Sciences

Macromolecular X-ray Crystallography and Automated Biological Calorimetry Facilities Newsletter Summer 2015

Auto-ITC 200 gets a system upgrade VP-Cap DSC has a new heater Next free training sessions for auto ITC, DSC and CD is scheduled for Sept. 24, 2015

1.  The automated Isothermal titration calorimeter has just received an upgrade

The auto ITC200 has just received a system upgrade.

A full preventative maintenance was also conducted on the equipment and the instrument is performing solidly.  ITC is a thermodynamic technique that directly measures the heat released or absorbed during a biomolecular binding event. ITC provides accurate determination of binding affinity (Kd), stoichiometry (n), enthalpy (ΔH) and entropy (ΔS).

 

 

2.  VP-Cap Differential Scanning Calorimeter has a new heater

The cap-VP DSC has just gone through a complete overhaul at the factory.  The Peltier heater for the cells was replaced and both cells were thoroughly cleaned and recalibrated.  Users have remarked that the data looks sharper and the equipment is more sensitive than ever.  

 

 

3.  Don’t miss out on the next training opportunity at the Automated Biological Calorimetry Facility

The next free training session for auto ITC and DSC is (Sept. 24, 1pm-3pm).You will be instructed on proper sample preparation techniques, how to use the “Design of Experiments” feature in the software in order to achieve optimal concentrations, as well as how to set up your experiments using the 96 well tray on the fully automated equipment.  Contact Julia Fecko (jaf48@psu.edu) in order to sign up for the next session. CD spectroscopy training (Sept. 24 10:30am-12pm).

 

 

4.  Research Underway

Cap VP-DSC

  • Seung-Yi Lee (Cremer) is using DSC in order to investigate the effects of osmolytes on thermoresponsive model polypeptides. This thermoresponsive peptide polymer folds upon temperature increase and this phenomenon is analogous to cold denaturation. The model peptides are elastin-like proteins that consist of pentapeptide repeats which are much simpler than real proteins. Experiments were conducted using a V5 type which is not charged. Osmolytes used were TMAO, betaine and glycine that contains methyl groups and C-O functional group. The DSC was used to investigate whether these osmolytes would favorably interact with the polypeptides upon folding by increasing temperature and by measuring enthalpy change.
  • Declan Evans (Showalter) is using the DSC to determine an appropriate temperature range for his study of the interactions of TRBP with double stranded RNA.
  • Kate O’Rourke (Boehr) Used DSC to compare thermal profiles of wildtype and 3 other proteins, A71G, A198G and A216G
  • Adrienne Green (Sandia National Laboratories) Used DSC to confirm if there was any unfolding of microtubules, tubulin proteins and f-actin in the lower 30-40°C range.

 

Auto ITC200

  • Kate O’Rourke (Boehr) is using the ITC to study the binding of tryptophan synthase subunits. The Boehr lab have identified a long-range network of amino acids in the alpha subunit that, when perturbed by alanine to glycine substitutions, hinder catalysis. The ITC is being used to help understand how these small perturbations impacts the binding of the alpha to the beta subunits in an effort to eventually understand the impact of inter-subunit communication.

 

5.  Facility acknowledged in publications

  1. Spectrin tetramer formation is not required for viable development in Drosophila. Khanna MR, Mattie FJ, Browder KC, Radyk MD, Crilly SE, Bakerink KJ, Harper SL, Speicher DW, Thomas GH,(2015) J BiolChem,  290(2), 706-15.
  2. Efficient Delivery of Long-Chain Fatty Aldehydes from the Nostocpunctiforme Acyl–Acyl Carrier Protein Reductase to its Cognate Aldehyde Deformylating Oxygenase Warui D, Pandelia, ME, Rajakovich L, Krebs C, Bollinger JM, Booker S, (2015) Biochemistry. 54(4): 1006-15. 
  3. Crystal structure of the PRC1 ubiquitylation module bound to the nucleosome. McGinty RK, Henrici RC, Tan S. (2014) Nature, 514(7524), 591-6
  4. Thermodynamic and structural determinants of differential Pdx1 binding to elements from the insulin and IAPP promoters. Bastidas M, Showalter SA. (2013) J Mol Biol., 425(18), 3360-77

 

Recent co-authored publications

  1. Crystallography:Precise Manipulation and Patterning of Protein Crystals for Macromolecular Crystallography Using Surface Acoustic Waves  Guo F, Zhou W, Li P, Mao Z, Yennawar NH, French JB, Huang TJ, (2015) Small. 11(23), 2710

 

  1. Structural, Electronic, and Magnetic Characterization of a Dinuclear Zinc Complex Containing TCNQ(-) and a µ-[TCNQ-TCNQ](2-) Ligand Kim J, Silakov A, Yennawar HP, Lear BJ, (2015) Inorg Chem 54(13), 6072-4.

 

  1. Structural models of mammalian mitochondrial transcription factor B2 Moustafa IM, Uchida A, Wang Y, Yennawar N, Cameron CE, (2015) Biochem Biophys Acta. 1849(8), 987-1002

 

  1. Crystal structures of 2-(4-nitro-phen-yl)-3-phenyl-2,3-di-hydro-4H-1,3-benzo-thia-zin-4-one and 2-(2-nitro-phen-yl)-3-phenyl-2,3-di-hydro-4H-1,3-benzo-thia-zin-4-one  Yennawar H, Cali AS, Xie Y, Silverberg LJ, (2015) Acta Crystallogr E Crystallogr Commun Mar 25;71(Pt 4):414-7

 

  1. Structural and Biochemical Characterization of a Ferredoxin:Thioredoxin Reductase-like Enzyme from Methanosarcina acetivorans  Kumar AK, Kumar RS, Yennawar NH, Yennawar HP, Ferry JG.(2015) Biochemistry 54(19), 3122-8

 

  1. Crystal structures of 2,3-bis-(4-chloro-phen-yl)-1,3-thia-zolidin-4-one and trans-2,3-bis-(4-chloro-phen-yl)-1,3-thia-zolidin-4-one 1-oxide  Yennawar HP, Tierney J, Hullihen PD, Silverberg LJ.(2015) Acta Crystallogr E Crystallogr Commun  Feb 11;71(Pt 3):264-7

 

  1. Crystal structure of N-[(2S,5R)-4-oxo-2,3-diphenyl-1,3-thia-zinan-5-yl]acetamide 0.375-hydrate  Yennawar HP, Singh H, Silverberg LJ. Acta Crystallogr E Crystallogr Commun. (2015) Jan 1;71(Pt 1):62-4

 

  1. Precise Manipulation and Patterning of Protein Crystals for Macromolecular Crystallography Using Surface Acoustic Waves. Guo F, Zhou W, Li P, Mao Z, Yennawar NH, French JB, Huang TJ, (2015) Small. 11(23), 2733-7

 

  1. Reversible intermolecular e-h oxidative addition to a geometrically deformed and structurally dynamic phosphorous triamide.Zhao W, McCarthy SM, Lai TY, Yennawar HP, Radosevich AT.,(2014) J Am Chem Soc., 136(50), 17634-44

 

  1. 2,3-Diphenyl-1,3-thia-zolidin-4-one.Yennawar HP, Tierney J, Silverberg LJ.,(2014) ActaCrystallogr Sect E Struct Rep Online. Jul 2;70(Pt 8):o847

 

  1. 2,3-Diphenyl-2,3-di-hydro-4H-pyrido[3,2-e][1,3]thia-zin-4-one.Yennawar HP, Singh H, Silverberg LJ.,(2014) ActaCrystallogr Sect E Struct Rep Online. May 3;70(Pt 6):o638

 

  1. 2,3-Diphenyl-2,3-di-hydro-4H-1,3-benzo-thia-zin-4-one.Yennawar HP, Bendinsky RV, Coyle DJ, Cali AS, Silverberg LJ., (2014) ActaCrystallogr Sect E Struct Rep Online. Mar 22;70(Pt 4):o465

 

  1. 2,3-Diphenyl-2,3,5,6-tetra-hydro-4H-1,3-thia-zin-4-one.Yennawar HP, Silverberg LJ., (2014) Acta Crystallogr Sect E Struct Rep Online. Jan 15;70(Pt 2):o133

 

  1. Autoinhibition of the Ron receptor tyrosine kinase by the juxtamembrane domain. Wang X, Yennawar N, Hankey PA., (2014) Cell Commun Signal., doi:10.1186/1478-811X-12-28

 

  1. Intermolecular N-H oxidative addition of ammonia, alkylamines, and arylamines to a planar σ(3)-phosphorus compound via an entropy-controlled electrophilic mechanism. McCarthy SM, Lin YC, Devarajan D, Chang JW, Yennawar HP, Rioux RM, Ess DH, Radosevich AT. (2014) J Am Chem Soc., 136(12), 4640-50
 

6.  Contact us

If you would like to sign up for free user-training on ITC, DSC, CD or Phoenix crystallography robot, contact director Neela Yennawar, PhD nhy1@psu.edu 814-863-9387 or research technician, Julia Fecko jaf48@psu.edu 814-865-8068. We will be happy to schedule free hands-on training.