Lawrence W. Miller

Associate Professor

Website: Miller's Research Group

Education:

B.S. 1989, The University of Wisconsin-Madison; Ph.D. 1998, The University of Wisconsin-Madison; Postdoctoral/Associate Research Scientist 2001-2006, Columbia University

The Miller laboratory develops and applies chemical methods to label proteins and study their function in vitro and in living mammalian cells. With chemical labeling, a protein of interest is expressed as a fusion to a polypeptide tag that interacts specifically with a synthetic fluorophore or other chemical probe. Our experimental approach incorporates synthetic organic chemistry, molecular and cell biology, and microscopy. Specific projects include:
Development of protein-targeted lanthanide luminophores.
Lanthanide-based resonance energy transfer (LRET) imaging of protein-protein interactions in live cells.
Time-gated luminescence microscopy.
LRET-based high-thoughput screening of protein interactions and their inhibition.
Cell penetrating peptide (CPP)-mediated delivery of imaging probes.

miller_fig_1
Figure 1. Selective labeling of a fusion protein with a small molecule in living cells. Epi-fluorescence micrographs (left, bright field; right, fluorescence) of NIH3T3 cells that were transiently transfected with DNA encoding a fusion of myosin light chain (MLC) to E. coli dihydrofolate reductase (eDHFR). The cells were incubated in growth medium containing a heterodimeric conjugate of trimethoprim (TMP) covalently linked to a fluorescein derivative. The TMP-fluorescein diffuses into cells and binds selectively and non-covalently to the MLC-eDHFR fusion protein which is associated with actin stress fibers. Selective chemical labeling can be used to impart fluorescence or other functionality to proteins in living cells for microscopy, screening and other biological analyses.

miller_fig_2
Figure 2. (upper left) Principle of time-resolved detection. Short pulses of illumination excite samples. Lanthanide probe molecules with long luminescent lifetimes (millisecond) are detected after a brief (microsecond) delay, eliminating short-lifetime scattering and autofluorescence background signals. We have developed a time-resolved epi-fluorescence microscope to image lanthanide luminescence in living cells at high signal-to-background ratio. (lower right) A luminescent terbium protein label. Probes of this type selectively impart lanthanide luminescence to fusion proteins, allowing time-resolved imaging and detection in vitro and in living cells.


SELECTED PUBLICATIONS


Mohandessi, S.; Rajendran, M.; Magda, D.; Miller L.W. (2012) ”Cell-penetrating peptides as delivery vehicles for a protein-targeted terbium complex.” Chem. Eur. J.
DOI:10.1002/chem.201201805

Yapici, E.; Reddy, D.R.; Miller L.W. (2012) “An adaptable luminescence resonance energy transfer assay for measuring and screening protein–protein interactions and their inhibition.” Chembiochem, 13, 553-558. (Featured Cover Article)

Rajapakse H.E.; Gahlaut N.; Mohandessi S.; Yu D.; Turner J.R.; Miller L.W. (2010) “Time-resolved luminescence resonance energy transfer imaging of protein–protein interactions in living cells.” Proc. Natl. Acad. Sci. USA, 107, 13582-13587.

Gahlaut N.; Miller L.W. (2010) “Time-resolved microscopy for imaging lanthanide luminescence in living cells.” Cytometry Part A, 77A, 1113-1125
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Contact Information

Office: 5205 SES, MC 111
Phone: 312-996-8542
Email: lwm2006@uic.edu