Carol Deutsch

Professor

215-898-8014
cjd@pennmedicine.upenn.edu

650A Clinical Research Building

415 Curie Boulevard

Philadelphia, PA 19104

Research Description

Our program aims to elucidate fundamental principles of protein folding in the formation of a voltage-gated potassium (Kv) channel, starting with its inception at the ribosome where Kv mRNA is translated and proceeding to the sequence of events underlying its integration into the bilayer of the endoplasmic reticulum. Knowledge of the mature Kv channel in the plasma membrane cannot tell us about the channel’s history of distinct biogenic stages; thus we focus on early folding events of the Kv protein. The stepwise progression of Kv channel formation is governed by a dynamic interaction between Kv protein, the ribosomal exit tunnel, and the translocon. The latter two are participatory conduits for nascent peptides during their synthesis and membrane integration. We use novel applications of our recently developed folding and accessibility assays, new unnatural amino acids/synthetase inhibitors, real-time translation kinetics, electrophysiology of Xenopus oocytes, photocrosslinking in the ribosomal tunnel and translocon, and computational approaches to probe the three major domains of a Kv channel (T1, voltage-sensor, and pore) and to define peptide-tunnel interactions for a broad range of peptides. Our findings will be particularly relevant to understanding defects in protein biogenesis and potential therapeutic strategies at the ribosome.

Data Protocols & Available Constructs

Tape Measure Constructs:

  • Extended tape measure nascent chains (single-cysteine mutants)
  • Poly-Arg constructs with T1 Kv1.3 N-terminus (WT and mutants)
  • Extended nascent chain with cysteine reporter and charged amino acids at the ribosome constriction
  • Poly-Ala constructs in an extended tape measure
  • Tandem tape measure constructs with/without poly-Ala tracts
  • Extended tape measures with folded N-terminal domains
  • Extended tape measures with truncated T1 domains
  • Poly-Ser constructs in an extended tape measure

Secondary Folding Assay:

  • Pegylation Protocols and cysteine-scanning constructs
  • Protocols using inserted poly-Ala tracks

Tertiary Folding Assay:

  • Pfold determination using bifunctional maleimides and PEG-MAL/PEG-SH
  • Modification kinetics using pegylation (kmod)

Location in the ribosomal exit tunnel:

  • Pegylation accessibility assay
  • Photocrosslinking using nitrene-generator
  • TMA modification kinetics (kmod)

Electrostatic Potential Determinations:

  • Constructs with reporter cysteines, poly-Arg and poly-Asp tracts

Unnatural Amino Acid Incorporation:

  • Flexizyme protocols for aminoacylation of tRNAs
  • Synthetase inhibition protocols
  • Rescue of arrested peptides at Cysteine or Amber codons

Diversity & Inclusion Initiatives

  • Mentor and Lecturer, Penn SUIP and PREP programs
  • Host of Workshop in Cell Biology for STEM undergrads from underrepresented communities
  • Member, Admissions committee for Cell Biology, Physiology and Metabolism (CPM) graduate group, which facilitates recruitment of diversity trainees. 

 

 

Degrees & Education

BA, Brandeis University, 1966

MPhil, Yale University, 1969

PhD, Yale University, 1972

Honors & Awards

2003-2007 NIH Study Section (MDCN-3)

2003-2004 President, Society of General Physiologists

2001-2002 Present Editorial Board, Journal of General Physiology

1996-1998 Society of General Physiologists Counci

1990 Lindback Distinguished Teaching Award, University of Pennsylvania

1985-1988 NSF Cell Physiology Advisory Panel

1981-1986 Research Career Development Award

Other Perelman School of Medicine Affiliations

Cell and Molecular Biology Graduate Program

Program in Cellular Physiology

Professional Affiliations

Society of General Physiologists

Biophysical Society

Recent Publications
August 3, 2020
The activation gate controls steady-state inactivation and recovery from inactivation in Shaker
Carol Deutsch, Ph.D.
Tibor Szanto, Florina Zakany, Ferenc Papp, Zoltan Varga, Carol Deutsch, & Gyorgy Panyi - DOI:10.1085/jgp.202012591 - PMID: 32442242 - PMCID: PMC7398138
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May 15, 2020
Forcing the ribosome to change its message
Carol Deutsch, Ph.D.
Sarah E. Leininger, Carol Deutsch, & Edward P. O’Brien - DOI: 10.1074/jbc.H120.013747 - PMCID: PMC7242706 - PMID: 32414911
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June 17, 2020
Side‐chain thioamides as fluorescence quenching probes
Carol Deutsch, Ph.D.
D. Miklos Robkis, Eileen M. Hoang, Pengse Po, Carol J. Deutsch, & E. James Petersson - PMID: 32740927 - DOI: 10.1002/bip.23384
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