Cargando…

Ligand-binding characteristics of feline insulin-binding immunoglobulin G

Polyclonal immunoglobulin (Ig) G autoantibodies against insulin have been identified in sera of healthy cats. We purified and fractionated insulin-binding IgGs from cat sera by affinity chromatography and analyzed affinity of insulin-binding IgGs for insulin and their epitopes. Following the passing...

Descripción completa

Detalles Bibliográficos
Autores principales: SUZUKI, Takafumi, NISHII, Naohito, TAKASHIMA, Satoshi, MATSUBARA, Tatsuya, IWASAWA, Atsushi, TAKEUCHI, Hirofumi, TAHARA, Kohei, HACHISU, Tatsuyuki, KITAGAWA, Hitoshi
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The Japanese Society of Veterinary Science 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4667653/
https://www.ncbi.nlm.nih.gov/pubmed/26062435
http://dx.doi.org/10.1292/jvms.15-0131
_version_ 1782403869552148480
author SUZUKI, Takafumi
NISHII, Naohito
TAKASHIMA, Satoshi
MATSUBARA, Tatsuya
IWASAWA, Atsushi
TAKEUCHI, Hirofumi
TAHARA, Kohei
HACHISU, Tatsuyuki
KITAGAWA, Hitoshi
author_facet SUZUKI, Takafumi
NISHII, Naohito
TAKASHIMA, Satoshi
MATSUBARA, Tatsuya
IWASAWA, Atsushi
TAKEUCHI, Hirofumi
TAHARA, Kohei
HACHISU, Tatsuyuki
KITAGAWA, Hitoshi
author_sort SUZUKI, Takafumi
collection PubMed
description Polyclonal immunoglobulin (Ig) G autoantibodies against insulin have been identified in sera of healthy cats. We purified and fractionated insulin-binding IgGs from cat sera by affinity chromatography and analyzed affinity of insulin-binding IgGs for insulin and their epitopes. Following the passing of fraction A, which did not bind to insulin, insulin-binding IgGs were eluted into two fractions, B and C, by affinity chromatography using a column fixed with bovine insulin. Dissociation constant (KD) values between insulin-binding IgGs and insulin, determined by surface plasmon resonance analysis (Biacore™system), were 1.64e(−4) M for fraction B (low affinity IgGs) and 2e(−5) M for fraction C (high affinity IgGs). Epitope analysis was conducted using 16 peptide fragments synthesized in concord with the amino acid sequence of feline insulin by an enzyme-linked immunosorbent assay. Fractions B and C showed higher absorbance (affinity) of the peptide fragment of 10 amino acid residues at the carboxyl-terminal of the B chain (peptide No. 19), followed by peptide fragments of 6 to 15 amino acid residues of the B chain (peptide No. 8). Fraction C showed a higher absorbance to 7 to 16 amino acid residues of the B chain (peptide No. 5) compared with the absorbance of fraction B. Polyclonal insulin-binding IgGs may form a macromolecule complex with insulin through the multiple affinity sites of IgG molecules. Feline insulin-binding IgGs are multifocal and may be composed of multiple IgG components and insulin.
format Online
Article
Text
id pubmed-4667653
institution National Center for Biotechnology Information
language English
publishDate 2015
publisher The Japanese Society of Veterinary Science
record_format MEDLINE/PubMed
spelling pubmed-46676532015-12-03 Ligand-binding characteristics of feline insulin-binding immunoglobulin G SUZUKI, Takafumi NISHII, Naohito TAKASHIMA, Satoshi MATSUBARA, Tatsuya IWASAWA, Atsushi TAKEUCHI, Hirofumi TAHARA, Kohei HACHISU, Tatsuyuki KITAGAWA, Hitoshi J Vet Med Sci Biochemistry Polyclonal immunoglobulin (Ig) G autoantibodies against insulin have been identified in sera of healthy cats. We purified and fractionated insulin-binding IgGs from cat sera by affinity chromatography and analyzed affinity of insulin-binding IgGs for insulin and their epitopes. Following the passing of fraction A, which did not bind to insulin, insulin-binding IgGs were eluted into two fractions, B and C, by affinity chromatography using a column fixed with bovine insulin. Dissociation constant (KD) values between insulin-binding IgGs and insulin, determined by surface plasmon resonance analysis (Biacore™system), were 1.64e(−4) M for fraction B (low affinity IgGs) and 2e(−5) M for fraction C (high affinity IgGs). Epitope analysis was conducted using 16 peptide fragments synthesized in concord with the amino acid sequence of feline insulin by an enzyme-linked immunosorbent assay. Fractions B and C showed higher absorbance (affinity) of the peptide fragment of 10 amino acid residues at the carboxyl-terminal of the B chain (peptide No. 19), followed by peptide fragments of 6 to 15 amino acid residues of the B chain (peptide No. 8). Fraction C showed a higher absorbance to 7 to 16 amino acid residues of the B chain (peptide No. 5) compared with the absorbance of fraction B. Polyclonal insulin-binding IgGs may form a macromolecule complex with insulin through the multiple affinity sites of IgG molecules. Feline insulin-binding IgGs are multifocal and may be composed of multiple IgG components and insulin. The Japanese Society of Veterinary Science 2015-06-09 2015-11 /pmc/articles/PMC4667653/ /pubmed/26062435 http://dx.doi.org/10.1292/jvms.15-0131 Text en ©2015 The Japanese Society of Veterinary Science http://creativecommons.org/licenses/by-nc-nd/3.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution Non-Commercial No Derivatives (by-nc-nd) License.
spellingShingle Biochemistry
SUZUKI, Takafumi
NISHII, Naohito
TAKASHIMA, Satoshi
MATSUBARA, Tatsuya
IWASAWA, Atsushi
TAKEUCHI, Hirofumi
TAHARA, Kohei
HACHISU, Tatsuyuki
KITAGAWA, Hitoshi
Ligand-binding characteristics of feline insulin-binding immunoglobulin G
title Ligand-binding characteristics of feline insulin-binding immunoglobulin G
title_full Ligand-binding characteristics of feline insulin-binding immunoglobulin G
title_fullStr Ligand-binding characteristics of feline insulin-binding immunoglobulin G
title_full_unstemmed Ligand-binding characteristics of feline insulin-binding immunoglobulin G
title_short Ligand-binding characteristics of feline insulin-binding immunoglobulin G
title_sort ligand-binding characteristics of feline insulin-binding immunoglobulin g
topic Biochemistry
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4667653/
https://www.ncbi.nlm.nih.gov/pubmed/26062435
http://dx.doi.org/10.1292/jvms.15-0131
work_keys_str_mv AT suzukitakafumi ligandbindingcharacteristicsoffelineinsulinbindingimmunoglobuling
AT nishiinaohito ligandbindingcharacteristicsoffelineinsulinbindingimmunoglobuling
AT takashimasatoshi ligandbindingcharacteristicsoffelineinsulinbindingimmunoglobuling
AT matsubaratatsuya ligandbindingcharacteristicsoffelineinsulinbindingimmunoglobuling
AT iwasawaatsushi ligandbindingcharacteristicsoffelineinsulinbindingimmunoglobuling
AT takeuchihirofumi ligandbindingcharacteristicsoffelineinsulinbindingimmunoglobuling
AT taharakohei ligandbindingcharacteristicsoffelineinsulinbindingimmunoglobuling
AT hachisutatsuyuki ligandbindingcharacteristicsoffelineinsulinbindingimmunoglobuling
AT kitagawahitoshi ligandbindingcharacteristicsoffelineinsulinbindingimmunoglobuling