Full metadata
Title
CHALLENGES IN THE EXPRESSION AND PURIFICATION OF INTERCELLULAR ADHESION MOLECULE- 1
Description
The Intercellular Adhesion Molecule-1 (ICAM-1, known as CD54) is a cell surface type I transmembrane glycoprotein with a molecular weight of 85 to 110 kDa. The primary function of ICAM-1 is to provide adhesion between endothelial cells and leukocytes after injury or stress. ICAM-1 is used as a receptor for various pathogens such as rhinoviruses, coxsackievirus A21 and the malaria parasite Plasmodium falciparum. ICAM-1 contains five immunoglobulin (Ig) domains in its long N-terminal extracellular region, a hydrophobic transmembrane domain, and a small C-terminal cytoplasmic domain. The Ig domains 1-2 and Ig domains 3-4-5 have been crystallized separately and their structure solved, however the full ICAM-1 structure has not been solved. Because ICAM-1 appears to be important for the mediation of cell-to-cell communication in physiological and pathological conditions, gaining a structural understanding of the full-length membrane anchored ICAM-1 is desirable. In this context, we have transiently expressed a plant-optimized gene encoding human ICAM-1 in Nicotiana benthamiana plants using the MagnICON expression system. The plant produced ICAM-1 is forming aggregates according to previous data. Thus, the current extraction and purification protocols have been altered to include TCEP, a reducing agent. The protein was purified using TALON metal affinity resin and partially characterized using various biochemical techniques. Our results show that there is a reduction in aggregation formation with the use of TCEP.
Date Created
2015-05
Contributors
- Patel, Heeral (Author)
- Mor, Tsafrir (Thesis director)
- Mason, Hugh (Committee member)
- Kannan, Latha (Committee member)
- Barrett, The Honors College (Contributor)
- School of Life Sciences (Contributor)
Topical Subject
Resource Type
Extent
14 pages
Language
eng
Copyright Statement
In Copyright
Primary Member of
Series
Academic Year 2014-2015
Handle
https://hdl.handle.net/2286/R.I.29037
Level of coding
minimal
Cataloging Standards
System Created
- 2017-10-30 02:50:57
System Modified
- 2021-08-11 04:09:57
- 3 years 3 months ago
Additional Formats