Abstract

Using a small volume mixer with the """"""""continuous flow"""""""" method, the flow speed can be made very high, which accelerates the generation of turbulence in the sample flow and thus promotes efficient mixing. Dr. Denis Rousseau has developed such a mixer for resonance Raman and fluorescence spectroscopies, and we have modified the design for use with the synchrotron infrared beam. The mixer is designed such that two solutions are loaded into syringes and pumped at 2 mm/ms into a small mixing well (100 ?m wide x 100 ?m deep) etched into a stainless steel block. The mixing well is covered by a 50 ?m-thick teflon sheet, which has a 200 ?m-diameter hole aligned in the center of the mixing well. The solutions mix in the mixing well and flow through the teflon spacer into the infrared """"""""sandwich"""""""" cell. The cell is made of two, rectangular CaF2 windows (12 mm x 25 mm), separated by a mylar Hspacer (100 ?m thick). The spacer creates a channel 200 ?m wide Hthrough the cell and provides a suitable infrared pathlength. By Hmoving the position of the infrared beam along the length of the infrared cell, different time points in the folding reaction can be probed. Since the dead time of the mixer is ~100 ?s, the time resolution of this rapid-mix flow cell is on the order of hundreds of microseconds to a few milliseconds.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Center for Research Resources (NCRR)
Type
Biotechnology Resource Grants (P41)
Project #
5P41RR001633-20
Application #
6618369
Study Section
Project Start
2002-09-01
Project End
2003-08-31
Budget Start
Budget End
Support Year
20
Fiscal Year
2002
Total Cost
Indirect Cost

  Institution

Name
Albert Einstein College of Medicine
Department
Type
DUNS #
009095365
City
Bronx
State
NY
Country
United States
Zip Code
10461

  Publications

Vongsvivut, Jitraporn; Fernandez, Jason; Ekgasit, Sanong et al. (2004) Characterization of supported cylinder-planar germanium waveguide sensors with synchrotron infrared radiation. Appl Spectrosc 58:143-51
Masip, Lluis; Pan, Jonathan L; Haldar, Suranjana et al. (2004) An engineered pathway for the formation of protein disulfide bonds. Science 303:1185-9
Huang, Raymond Y; Miller, Lisa M; Carlson, Cathy S et al. (2003) In situ chemistry of osteoporosis revealed by synchrotron infrared microspectroscopy. Bone 33:514-21
Rashidzadeh, Hassan; Khrapunov, Sergei; Chance, Mark R et al. (2003) Solution structure and interdomain interactions of the Saccharomyces cerevisiae ""TATA binding protein"" (TBP) probed by radiolytic protein footprinting. Biochemistry 42:3655-65
Uchida, Takeshi; Takamoto, Keiji; He, Qin et al. (2003) Multiple monovalent ion-dependent pathways for the folding of the L-21 Tetrahymena thermophila ribozyme. J Mol Biol 328:463-78
Taylor, Colleen M; Watton, Stephen P; Bryngelson, Peter A et al. (2003) Inner-sphere complexation of cobalt(II) 2,9-dimethyl-1,10-phenanthroline ([Co(neo)]2+) with commercial and sol-gel derived silica gel surfaces. Inorg Chem 42:312-20
Dhavan, Gauri M; Crothers, Donald M; Chance, Mark R et al. (2002) Concerted binding and bending of DNA by Escherichia coli integration host factor. J Mol Biol 315:1027-37
Uchida, Takeshi; He, Qin; Ralston, Corie Y et al. (2002) Linkage of monovalent and divalent ion binding in the folding of the P4-P6 domain of the Tetrahymena ribozyme. Biochemistry 41:5799-806
Tang, Qun; Carrington, Paul E; Horng, Yih-Chern et al. (2002) X-ray absorption and resonance Raman studies of methyl-coenzyme M reductase indicating that ligand exchange and macrocycle reduction accompany reductive activation. J Am Chem Soc 124:13242-56
Guan, Jing-Qu; Vorobiev, Sergeui; Almo, Steven C et al. (2002) Mapping the G-actin binding surface of cofilin using synchrotron protein footprinting. Biochemistry 41:5765-75

Showing the most recent 10 out of 68 publications