Difference between revisions of "Part:BBa K3260024"

Revision as of 14:50, 29 July 2020

Teriparatide with stop codon

Teriparatide is the first 34 amino acids of Parathyroid hormone and is used to treat osteoporosis.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

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 <!-- Add more about the biology of this part here
 ===Usage and Biology===
+Our group used a structural approach to measure the concentrations of PTH in the interstitial fluid. This may improve this teams part of providing 1-34 PTH for therapeutics. With a PTH protein based assay, clinicians may make better decisions regarding dosing of PTH and treatment. A recent study did assess the use of PTHR and FRET to measure the concentrations of PTH. This method did not go as far as to include the detection of PTH in a biosensor. Using the 60 amino acid N- terminal, extracellular domain, of PTHR to bind PTH, this would allow for easier protein expression. We added a mNeonGreen fluorophore to the N-termini of this extracellular domain. This domain had one cysteine and so adding our E/K coiled coil system that immobilizes the protein was not a large concern.
+The issue at hand is rather detecting the concentration of PTH in the body. While PTH is difficult to express, a PTH fragment (residues 1-34) is easier to express (or obtain). This PTH fragment is attached to a mCherry fluorophore through its one lysine residues amino group. Allowing the PTH fragment with mCherry and PTHR with mNeonGreen to associate produces a FRET signal. When PTH from the body enters the biosensor, it displaces the PTH with mCherry and a drop in FRET signal is observed. You may consider this as a competition assay or similar to an antibody detection system.
+Nie Y, Li J, Liu Y, Zhang Q, Ma Q. A Visual FRET Immunofluorescent Biosensor for Ratiometric Parathyroid Hormone (1-84) Antigen Point-of-Care Detection. J Fluoresc. 2020 Mar;30(2):329–34.
+[[Image:pthcompetitiveassay.png|800px]]
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