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The structure-activity relationship study of the N-terminal domain in desert locust ion transport peptide (ITP) Zhao, Ying
Abstract
The ileum and rectum of locust hindgut constitute the reabsorptive part of the excretory system. They are functionally analogous to a proximal and distal parts of convoluted tubule of the mammalian kidney tubule, respectively. Ion transport peptide (ITP) purified from locust nervous corpus cardiacum (CC) has been shown to stimulate salt and water reabsorption and inhibit acid secretion in the ileum of Schistocerca gregaria . The primary structure of ITP deduced from its cDNA suggests that it is a 72 amino acid peptide with C-terminal amidation and three disulfide bonds. Both synthetic and expressed ITP mimic the biological activity of ITP purified from S.gregaria CC. It has been demonstrated that ITP is a true member of the CHH (Crustacean Hyperglycemic Hormone) family. This study examines the structure-activity relationship of the N-terminal domain of ITP. Two questions are addressed: 1. Is the N-terminal domain of ITP consisting of the first six amino acids (SFFDIQ) important to bioactivity? 2. Which amino acids in the N-terminal domain of ITP are essential for ITP binding to the receptor and/or activating the receptor? Using site-directed mutagenesis and voltage-clamped locust ileum as bioassay, I found the ITP N-terminal domain (SFFDIQ) is important for its bioactivity. Among the six amino acid of ITP N-terminus, Phe2 and Phe3 are essential for ITP binding to the receptor, and Phe2 is also important to receptor activation. The other four amino acids SI, D4,15, and Q6 didn't contribute to the ITP bioactivity, even D4 is a highly conserved amino acid. Post-translational modification of conversion L- to D- amino acid probably occurs at Phe2 and Phe3 to yield two ITP isomorphs. Mutations on the ITP N-terminal domain didn't interfere with the dibasic cleavage site in spite of its immediate proximity to the dibasic cleavage site. Mutant F2A has the potential to be ITP antagonist.
Item Metadata
Title |
The structure-activity relationship study of the N-terminal domain in desert locust ion transport peptide (ITP)
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Creator | |
Publisher |
University of British Columbia
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Date Issued |
2000
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Description |
The ileum and rectum of locust hindgut constitute the reabsorptive part of the
excretory system. They are functionally analogous to a proximal and distal parts of
convoluted tubule of the mammalian kidney tubule, respectively. Ion transport peptide
(ITP) purified from locust nervous corpus cardiacum (CC) has been shown to stimulate salt
and water reabsorption and inhibit acid secretion in the ileum of Schistocerca gregaria . The
primary structure of ITP deduced from its cDNA suggests that it is a 72 amino acid peptide
with C-terminal amidation and three disulfide bonds. Both synthetic and expressed ITP
mimic the biological activity of ITP purified from S.gregaria CC. It has been demonstrated
that ITP is a true member of the CHH (Crustacean Hyperglycemic Hormone) family.
This study examines the structure-activity relationship of the N-terminal domain of
ITP. Two questions are addressed:
1. Is the N-terminal domain of ITP consisting of the first six amino acids (SFFDIQ)
important to bioactivity?
2. Which amino acids in the N-terminal domain of ITP are essential for ITP binding to
the receptor and/or activating the receptor?
Using site-directed mutagenesis and voltage-clamped locust ileum as bioassay, I found
the ITP N-terminal domain (SFFDIQ) is important for its bioactivity. Among the six amino acid
of ITP N-terminus, Phe2 and Phe3 are essential for ITP binding to the receptor, and Phe2 is also
important to receptor activation. The other four amino acids SI, D4,15, and Q6 didn't contribute
to the ITP bioactivity, even D4 is a highly conserved amino acid. Post-translational modification
of conversion L- to D- amino acid probably occurs at Phe2 and Phe3 to yield two ITP
isomorphs. Mutations on the ITP N-terminal domain didn't interfere with the dibasic cleavage
site in spite of its immediate proximity to the dibasic cleavage site. Mutant F2A has the potential
to be ITP antagonist.
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Extent |
4847883 bytes
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Genre | |
Type | |
File Format |
application/pdf
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Language |
eng
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Date Available |
2009-07-09
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Provider |
Vancouver : University of British Columbia Library
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Rights |
For non-commercial purposes only, such as research, private study and education. Additional conditions apply, see Terms of Use https://open.library.ubc.ca/terms_of_use.
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DOI |
10.14288/1.0089504
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URI | |
Degree | |
Program | |
Affiliation | |
Degree Grantor |
University of British Columbia
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Graduation Date |
2000-05
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Campus | |
Scholarly Level |
Graduate
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Aggregated Source Repository |
DSpace
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Item Media
Item Citations and Data
Rights
For non-commercial purposes only, such as research, private study and education. Additional conditions apply, see Terms of Use https://open.library.ubc.ca/terms_of_use.