Full metadata
Title
Expression of Pleiotrophin as Separate Domains to Examine Glycosaminoglycan Binding Sites
Description
Glycosaminoglycan (GAG) binding by the cytokine pleiotrophin (PTN) was examined by expressing both thrombospondin 1 type-1 repeat domains of PTN separately, as PTN-N and PTN-C. PTN-N contains residues 31-89, and PTN-C contains residues 90-146. Nuclear magnetic resonance (NMR) experiments were conducted on both PTN-N and PTN-C to elucidate GAG binding regions. Titration with heparin dp6 showed a twofold increase in affinity when expressing PTN-N and PTN-C separately rather than as intact PTN. Paramagnetic relaxation rate enhancement experiments and surface paramagnetic relaxation enhancement (PRE) perturbation experiments were used to determine which residues were involved in GAG binding. One binding site was observed in PTN-N, around residue T82, and two binding sites were observed in PTN-C, one around residue K93 and the other around residue G142. These observed binding sites agree with the binding sites already proposed by the Wang lab group and other studies. Future work on the subject could be done on confirming that other varieties and length GAGs bind at the same sites, as well as examining the effect longer GAG fragments have on the affinity of intact PTN versus separate domains.
Date Created
2015-12
Contributors
- Kuch, Nathaniel Jacob (Author)
- Wang, Xu (Thesis director)
- Van Horn, Wade (Committee member)
- School of Molecular Sciences (Contributor)
- School of Life Sciences (Contributor)
- Barrett, The Honors College (Contributor)
Topical Subject
Resource Type
Extent
28 pages
Language
eng
Copyright Statement
In Copyright
Primary Member of
Series
Academic Year 2015-2016
Handle
https://hdl.handle.net/2286/R.I.35833
Level of coding
minimal
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System Created
- 2017-10-30 02:50:57
System Modified
- 2021-08-11 04:09:57
- 3 years 3 months ago
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