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Baculovirus Expression and Purification of Wild Type and Mutant Full-Length Human LRRK2

Wang, Wen
http://rave.ohiolink.edu/etdc/view?acc_num=case1216849751

Abstract Details

2008, Master of Sciences, Case Western Reserve University, Environmental Health Science.
LRRK2 is a complex 285 kD protein in which the most prevailing mutations G2019S and R1441C are linked to autosomal dominant Parkinson disease (PD). This study has established an expression and purification system to produce active recombinant proteins of the LRRK2, to examine whether the mutations alter the LRRK2 activity and what mechanism underlies the alteration. Both bacterial and baculovirus expression systems were attempted to express and purify the proteins. The active full-length LRRK2 proteins (wild type, G2019S and R1441C) were obtained via the baculovirus system. The bacterial expression system was used for expression and purification of the LRRK2 kinase domain (LRRK2-KD) (wild type and G2019S) but it displayed no activity. The purified recombinant proteins obtained via the baculovirus expression system were analyzed in kinase assays. Our preliminary estimates of Km and Vmax may provide insight regarding the mechanism of LRRK2 activation by PD-linked mutations.
Martina L. Veigl, PhD (Committee Chair)
Qingzhong Kong, PhD (Advisor)
Kumar Alagramam, PhD (Committee Member)
John J. Mieyal, PhD (Committee Member)
David G McCoy, PhD (Committee Member)
92 p.
Molecular Biology
PD; LRRK2; Baculovirus expression systems

Recommended Citations

Citations

  • Wang, W. (2008). Baculovirus Expression and Purification of Wild Type and Mutant Full-Length Human LRRK2 [Master's thesis, Case Western Reserve University]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=case1216849751

    APA Style (7th edition)

  • Wang, Wen. Baculovirus Expression and Purification of Wild Type and Mutant Full-Length Human LRRK2. 2008. Case Western Reserve University, Master's thesis. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=case1216849751.

    MLA Style (8th edition)

  • Wang, Wen. "Baculovirus Expression and Purification of Wild Type and Mutant Full-Length Human LRRK2." Master's thesis, Case Western Reserve University, 2008. http://rave.ohiolink.edu/etdc/view?acc_num=case1216849751

    Chicago Manual of Style (17th edition)

case1216849751
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© 2008, all rights reserved.
This open access ETD is published by Case Western Reserve University School of Graduate Studies and OhioLINK.