Identification and Characterization of Internalization Signal of the Prostate Specific Membrane Antigen
Abstract
The purpose of this award is to identify and characterize an internalization signal in the cytoplasmic tail of prostate specific membrane antigen PSMA, transmembrane protein abundantly expressed in prostate cancer cells. These studies should aid in understanding the mechanism of antibody uptake and fate of the internalized antibody to improve antibody delivery approaches for immunotherapy for prostate cancer. We have now established and standardized a culture model to study internalization signal of PSMA. In addition we have determined that the cytoplasmic tail of PSMA has an internalization signal. An alanine scan mutagenesis approach in which each of the amino acids in the cytoplasmic tail is mutated to an alanine indicates that a di-leucine motif in the cytoplasmic tail of PSMA may be involved in its internalization. Furthermore, the effect of specific amino acid mutation in targeting PSMA through the endocytic pathway is being tested by immunofluorescence and laser-scanning confocal microscopy. A cell-surface biotinylation assay is being utilized to quantify the internalization of PSMA cytoplasmic tail deletion and point mutants.
Document Details
- Document Type
- Technical Report
- Publication Date
- Aug 01, 1999
- Accession Number
- ADB253500
Entities
People
- Ayyappan K. Rajasekaran
Organizations
- University of California, Los Angeles