Biophysical Interaction and Characterization Facility (BICF)

Biophysical Interaction and Characterization Facility (BICF)

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Analytical Ultracentrifuge

Beckman Coulter ProteomeLab XL-I with An-50 Ti Rotor

Measure MW of Proteins, Formation of Dimers/Trimers/Etc., Association Constants

  • Equipped with both Absorbance and Interference Optics
  • Absorbance Optical Range: 190 – 800 nm; linear to 1.5 OD
  • Interference Optical Wavelength: 660 nm
  • Temperature Range: 0oC to 40oC
  • Max Speed: 50,000 rpm
  • 8-hole rotor (7 sample cells with counterbalance)

 

Circular Dichroism Spectrometer

Jasco J-815

Measure Protein/Peptide Secondary Structures, Chiral Organics

  • Wavelength Range: 165 – 1100 nm
  • 6-Cell Sample Holder for Automated Measurements (simultaneous spectra, parallel thermal melts, etc.)
    • Compatible with both 10 mm and 1 mm cuvettes (adapter provided)
  • Peltier Device Sample Holder for Thermal Scans
    • Temperature Range: 2-100°C
    • Each of the 6 sample holders controlled individually, can operate simultaneously with different thermal parameters

 

Isothermal Titration Calorimeter

MicroCal VP-ITC MicroCalorimeter

Measure Thermodynamic Parameters, Binding Affinity of Macromolecule Interactions

  • Working Sample Volume: 1.4 mL (~ 2 mL needed for proper injection without air pocket)
  • Ligand Injection Volume Range: 0.1-279.7 µL
  • Number of Injections: Single Injection Mode up to 500 Consecutive Injections
  • Temperature Range: 2-80 °C
  • Ni-Cr-Mo Alloy Sample Cell: highly resistant to basic solutions, buffers, chlorides
  • Adjustable Stirring Speed

 

Differential Scanning Calorimeter

MicroCal VP-DSC MicroCalorimeter

Measure Thermodynamics of Phase Transitions in Biomacromolecules/Polymers

  • Analyzes Samples in Solution Only – NO SOLIDS
  • Temperature Range: -10-130 °C
  • Working Sample Volume: 585 µL (750 µL needed for proper injection without air pocket)
  • Minimum Response Time: 7 seconds
  • Capable of Upscan and Downscan Measurements
    • Upscan Rates: 0 to 90°C per hour
    • Downscan Rates: 0 to -60°C per hour
  • Tantalum Alloy (Tantaloy 61™) Sample Cell

 

Stopped-Flow Spectrometer

Applied Photophysics SX-20

Measure kinetics of interactions

  • 20 µL Cell Volume with Dead Time of 1 ms
  • Capable of Absorbance and Fluorescence Measurements
    • Absorbance: 185-900 nm
    • Fluorescence (Em): 300-650 nm
  • Monochromator Slit Widths: 0.5-37 nm
  • Xe Lamp Source
  • Single-Mixing or Sequential-Mixing Modes Available
  • Water Circulator Accessory for Controlled Temperature Experiments
  • Anaerobic Adapter for Experiments in N2 Environment

 

Other Equipment

  • UV/Vis Spectrophotometer 
  • BioImager
  • Plate Reader 
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Select Publications - Analytical Ultracentrifuge

Choudhary, D.; Narui, Y.; Neel, B.L.; Wimalasena, L.N.; Klanseck, C.F.; De-la-Torre, P.; Chen, C.; Araya-Secchi, R.; Tamilselvan, E.; Sotomayor, M. Structural determinants of protocadherin-15 mechanics and function in hearing and balance perception. Proc. Natl. Acad. Sci. U. S. A. 2020, 117, 24837-24848. DOI: 10.1073/pnas.1920444117.

Chen, L.; Tanimoto, A.; So, B.R.; Bakhtina, M.; Magliery, T.J.; Wysocki, V.H.; Musier-Forsyth, K. Stoichiometry of triple-sieve tRNA editing complex ensures fidelity of aminoacyl-tRNA formation. Nucleic Acids Res. 2019, 47, 929-940. DOI: 10.1093/nar/gky1153.

Danhart, E.M.; Bakhtina, M.; Cantara, W.A.; Kuzmishin, A.B.; Ma, X.; Sanford, B.L.; Vargas-Rodriguez, O.; Košutić, M.; Goto, Y.; Suga, H.; Nakanishi, K.; Micura, R.; Foster, M.P.; Musier-Forsyth, K. Conformational and chemical selection by a trans-acting editing domain. Proc. Nat. Acad. Sci. U. S. A. 2019, 114, E6774–E6783. DOI: 10.1073/pnas.1703925114.

Select Publications - Circular Dichroism

Creemer, C.; Kilic, H.; Lee, K.S.; Saracoglu, N.; Parquette, J.R. Light-controlled self-assembly of a dithienylethene bolaamphiphile in water. Dalton Trans. 2020, 49, 8846-8849. DOI: 10.1039/D0DT02001J.

Wen, J.; Liao, H.; Stachowski, K.; Hempfling, J.P.; Qian, Z.; Yuan, C.; Foster, M.P.; Pei, D. Rational design of cell-permeable cyclic peptides containing a D-Pro-L-Pro motif. Bioorg. Med. Chem. 2020, 28, 115711. DOI: 10.1016/j.bmc.2020.115711.

Goyal, V.D.; Sullivan, B.J.; Magliery, T.J. Phylogenetic spread of sequence data affects fitness of consensus enzymes: Insights from triosephosphate isomerase. Proteins: Struct., Funct., Bioinf. 2020, 88, 274-283. DOI: 10.1002/prot.25799.

Select Publications - Calorimetry

Sen, S.; Hendricks, A.L.; Cowan, J.A. Cluster exchange reactivity of [2Fe-2S]-bridged heterodimeric BOLA1-GLRX5. FEBS J. 2020. DOI: 10.1111/febs.15452.

Bong, D.; Miao, S.; Liang, Y.; Mao, J.; Marathe, I. Duplex stem replacement with bPNA+ triplex hybrid stems enables reporting on tertiary interactions of internal RNA domains. ChemRxiv. 2019, Preprint. DOI: 10.26434/chemrxiv.7583624.v2.

Sen, S.; Rao, B.; Wachnowsky, C.; Cowan, J.A. Cluster exchange reactivity of [2Fe-2S] cluster-bridged complexes of BOLA3 with monothiol glutaredoxins. Metallomics 2018, 10, 1282-1290. DOI: 10.1039/C8MT00128F.

Select Publications - Stopped-Flow 

Kisgeropoulos, E.C.; Griese, J.J.; Smith, Z.R.; Branca, R.M.M.; Schneider, C.R.; Högbom, M.; Shafaat, H.S. Key Structural Motifs Balance Metal Binding and Oxidative Reactivity in a Heterobimetallic Mn/Fe Protein. J. Am. Chem. Soc. 2020, 142, 5338-5354. DOI: 10.1021/jacs.0c00333.

Coldren, W.H.; Tikunova, S.B.; Davis, J.P.; Lindert, S. Discovery of Novel Small-Molecule Calcium Sensitizers for Cardiac Troponin C: A Combined Virtual and Experimental Screening Approach. J. Chem. Inf. Model 2020, 60, 3648-3661. DOI: 10.1021/acs.jcim.0c00452.

Miller, E.K.; Trivelas, N.E.; Maugeri, P.T.; Blaesi, E.J.; Shafaat, H.S. Time-Resolved Investigations of Heterobimetallic Cofactor Assembly in R2lox Reveal Distinct Mn/Fe Intermediates. ACS Biochem. 2017, 56, 3369-3379. DOI: 10.1021/acs.biochem.7b00403.

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Staff

staff

Dr. Alicia Friedman 

BICF Lab: MP 0043
Office: CBEC 085 
614-292-9646
friedman.270@osu.edu
u.osu.edu/bicf