Michael D. Lane

Portrait of MD/PhD student Michael LaneEmail: lane0280@umn.edu

Entering Class:
2009

Education:

University of Florida
Microbiology and Cell Science major
B.S., 2006

University of Minnesota
Biochemistry, Molecular Biology and Biophysics Graduate Program
Ph.D., 2016

Honors and Awards:

  • University of Minnesota Doctoral Dissertation Fellowship, 2014-2015
  • American Heart Association Research Fellowship, 2012-2017

MSTP Student Governance:

  • Student Advisory Committee, 2015-17

Thesis Advisor: Burckhard Seelig, Ph.D.

Thesis Research

Directed evolution of streptopain to inactivate superantigens

Streptococcus pyogenes is a pervasive bacterium capable of causing a wide range of human disease, from the simple “strep throat” to the terrifying “flesh eating disease” (necrotizing fasciitis). When introduced to the bloodstream, S. pyogenes can release extremely potent toxins, named “superantigens,” causing streptococcal toxic shock syndrome (STSS) and death within hours. Proteolytic neutralization of superantigens would prevent these deadly toxicities, but superantigens have shown natural resistance to proteolysis. Recently, streptopain (a cysteine protease produced by S. pyogenes) was shown to degrade SmeZ (the most potent superantigen) yet not SpeA (a superantigen linked to STSS and necrotizing fasciitis) despite over 50% sequence identity and a very similar fold. We will evolve streptopain variants capable of cleaving SpeA by using mRNA display technology. This entirely in vitro method allows the selection of an active enzyme from libraries of up to 1013 variants. We will clone and express SpeA and streptopain, determine the streptopain cleavage site of SmeZ, engineer selection primers displaying the SpeA target region homologous to the SmeZ cleavage site, build a variant streptopain library with random mutations incorporated at specificity conferring regions, and select for SpeA-cleaving ability. We plan to establish the streptopain protease as a platform from which we will develop an arsenal of superantigen-neutralizing proteases to prevent the deadly toxicities of STSS and necrotizing fasciitis as well as defend against the myriad of other diseases linked to superantigens.

Publications (pubmed)

Lane MD, Seelig B. Highly efficient recombinant production and purification of streptococcal cysteine protease streptopain with increased enzymatic activity. Protein Expr Purif. 2016 May;121:66-72. PMCID: PMC4803605

Lane MD, Seelig B. Advances in the directed evolution of proteins. Curr Opin Chem Biol. 2014 Oct;22:129-36. Review. PMCID: PMC4253873

For work prior to entering the MSTP

Nam HJ, Lane MD, Padron E, Gurda B, McKenna R, Kohlbrenner E, Aslanidi G, Byrne B, Muzyczka N, Zolotukhin S, Agbandje-McKenna M. Structure of adeno-associated virus serotype 8, a gene therapy vector. J Virol. 2007 Nov;81(22):12260-71. PMCID: PMC2168965

Lane MD, Nam HJ, Padron E, Gurda-Whitaker B, Kohlbrenner E, Aslanidi G, Byrne B, McKenna R, Muzyczka N, Zolotukhin S, Agbandje-McKenna M. Production, purification, crystallization and preliminary X-ray analysis of adeno-associated virus serotype 8. Acta Crystallogr Sect F Struct Biol Cryst Commun. 2005 Jun 1;61(Pt 6):558-61. PMCID: PMC1952340