Limiting RPE lipofuscin accumulation by harnessing enzyme-mediated degradation

Project: Research project

Project Details

Description

Limiting RPE lipofuscin accumulation by harnessing enzyme-mediated degradation Limiting RPE lipofuscin accumulation by harnessing enzyme-mediated degradation Limiting RPE lipofuscin accumulation by harnessing enzyme-mediated degradation Work in the laboratory of Dr. Bruce Rittmann at Arizona State University will include the expression of peroxidase enzymes in Pichia pastoris using recombinant DNA techniques, the objective being to attach mannose residues to glycosylation motifs. Binding of the mannose residues to mannose receptors on the cell surface will serve to initiate endosomal uptake and targeting to lysosomes. Lysates of peroxidase-expressing Pichia will also be used to text enzyme activity. Other studies will involve the cloning of carotenoid cleaving dioxygenases (CCDs), for instance from cyanobacterial strains. CCDs will also be amplified from environmental DNA using degenerate PCR. The CCDs will be expressed in E. coli and efforts will also be directed towards delivering the enzymes to RPE lysosomes using glycosylation-independent targeting via mannose-6-phosphate receptors.
StatusFinished
Effective start/end date12/15/099/30/13

Funding

  • Edward N. & Della L. Thome Memorial Foundation: $300,000.00

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