The Binding Associations of the Mannose 6-Phosphate/Insulin-Like Growth Factor 2 Receptor
Advisor Information
Jodi Kreiling
Location
Dr. C.C. and Mabel L. Criss Library
Presentation Type
Poster
Start Date
6-3-2015 2:00 PM
End Date
6-3-2015 3:30 PM
Abstract
The Man6P/IGFIIR (mannose 6-phosphate/insulin-like growth factor II receptor) is a cell membrane protein involved in cellular growth regulation and tumor suppression. This protein has been found to function optimally as a dimer, where two whole proteins work together to ensure full functionality. Dimerization seems to involve multiple regions of the protein receptor, and no specific dimerization domain has been discovered. In an effort to study the dimerization patterns of the Man6P/IGFIIR, variable tagged domains of the receptor (FLAG (F) or Myc (M)) were constructed and tested for dimerization capability and strength of association. Although the data obtained indicated that association of all triplet domains was possible and that identical triplet domains had the strongest dimerization contacts, those results were unable to be quantified due to heavy chain interference from the antibody used in the binding assays. The goal of this project was to attach a 6x-histidine tag (H) to the end of two triplet receptor constructs, 7-9M and 1-3M, in order to increase their molecular weight for better interpretation of the binding and disruption assays using the triplet receptors and also allow for the use of nickel ion in binding assays instead of an antibody.
The Binding Associations of the Mannose 6-Phosphate/Insulin-Like Growth Factor 2 Receptor
Dr. C.C. and Mabel L. Criss Library
The Man6P/IGFIIR (mannose 6-phosphate/insulin-like growth factor II receptor) is a cell membrane protein involved in cellular growth regulation and tumor suppression. This protein has been found to function optimally as a dimer, where two whole proteins work together to ensure full functionality. Dimerization seems to involve multiple regions of the protein receptor, and no specific dimerization domain has been discovered. In an effort to study the dimerization patterns of the Man6P/IGFIIR, variable tagged domains of the receptor (FLAG (F) or Myc (M)) were constructed and tested for dimerization capability and strength of association. Although the data obtained indicated that association of all triplet domains was possible and that identical triplet domains had the strongest dimerization contacts, those results were unable to be quantified due to heavy chain interference from the antibody used in the binding assays. The goal of this project was to attach a 6x-histidine tag (H) to the end of two triplet receptor constructs, 7-9M and 1-3M, in order to increase their molecular weight for better interpretation of the binding and disruption assays using the triplet receptors and also allow for the use of nickel ion in binding assays instead of an antibody.