Comparison of recombinant human haptocorrin expressed in human embryonic kidney cells and native haptocorrin.

Haptocorrin (HC) is a circulating corrinoid binding protein with unclear function. In contrast to transcobalamin, the other transport protein in blood, HC is heavily glycosylated and binds a variety of cobalamin (Cbl) analogues. HC is present not only in blood but also in various secretions like mil...

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Autores principales:	Evelyne Furger, Sergey N Fedosov, Dorte Launholt Lildballe, Robert Waibel, Roger Schibli, Ebba Nexo, Eliane Fischer
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2012
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/e746c69532c449408e881ca48d381e85
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spelling	oai:doaj.org-article:e746c69532c449408e881ca48d381e852021-11-18T07:17:24ZComparison of recombinant human haptocorrin expressed in human embryonic kidney cells and native haptocorrin.1932-620310.1371/journal.pone.0037421https://doaj.org/article/e746c69532c449408e881ca48d381e852012-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/22662153/?tool=EBIhttps://doaj.org/toc/1932-6203Haptocorrin (HC) is a circulating corrinoid binding protein with unclear function. In contrast to transcobalamin, the other transport protein in blood, HC is heavily glycosylated and binds a variety of cobalamin (Cbl) analogues. HC is present not only in blood but also in various secretions like milk, tears and saliva. No recombinant form of HC has been described so far. We report the expression of recombinant human HC (rhHC) in human embryonic kidney cells. We purified the protein with a yield of 6 mg (90 nmol) per litre of cell culture supernatant. The isolated rhHC behaved as native HC concerning its spectral properties and ability to recognize both Cbl and its baseless analogue cobinamide. Similar to native HC isolated from blood, rhHC bound to the asialoglycoprotein receptor only after removal of terminal sialic acid residues by treatment with neuraminidase. Interestingly, rhHC, that compared to native HC contains four excessive amino acids (…LVPR) at the C-terminus, showed subtle changes in the binding kinetics of Cbl, cobinamide and the fluorescent Cbl conjugate CBC. The recombinant protein has properties very similar to native HC and although showing slightly different ligand binding kinetics, rhHC is valuable for further biochemical and structural studies.Evelyne FurgerSergey N FedosovDorte Launholt LildballeRobert WaibelRoger SchibliEbba NexoEliane FischerPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 7, Iss 5, p e37421 (2012)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Evelyne Furger Sergey N Fedosov Dorte Launholt Lildballe Robert Waibel Roger Schibli Ebba Nexo Eliane Fischer Comparison of recombinant human haptocorrin expressed in human embryonic kidney cells and native haptocorrin.
description	Haptocorrin (HC) is a circulating corrinoid binding protein with unclear function. In contrast to transcobalamin, the other transport protein in blood, HC is heavily glycosylated and binds a variety of cobalamin (Cbl) analogues. HC is present not only in blood but also in various secretions like milk, tears and saliva. No recombinant form of HC has been described so far. We report the expression of recombinant human HC (rhHC) in human embryonic kidney cells. We purified the protein with a yield of 6 mg (90 nmol) per litre of cell culture supernatant. The isolated rhHC behaved as native HC concerning its spectral properties and ability to recognize both Cbl and its baseless analogue cobinamide. Similar to native HC isolated from blood, rhHC bound to the asialoglycoprotein receptor only after removal of terminal sialic acid residues by treatment with neuraminidase. Interestingly, rhHC, that compared to native HC contains four excessive amino acids (…LVPR) at the C-terminus, showed subtle changes in the binding kinetics of Cbl, cobinamide and the fluorescent Cbl conjugate CBC. The recombinant protein has properties very similar to native HC and although showing slightly different ligand binding kinetics, rhHC is valuable for further biochemical and structural studies.
format	article
author	Evelyne Furger Sergey N Fedosov Dorte Launholt Lildballe Robert Waibel Roger Schibli Ebba Nexo Eliane Fischer
author_facet	Evelyne Furger Sergey N Fedosov Dorte Launholt Lildballe Robert Waibel Roger Schibli Ebba Nexo Eliane Fischer
author_sort	Evelyne Furger
title	Comparison of recombinant human haptocorrin expressed in human embryonic kidney cells and native haptocorrin.
title_short	Comparison of recombinant human haptocorrin expressed in human embryonic kidney cells and native haptocorrin.
title_full	Comparison of recombinant human haptocorrin expressed in human embryonic kidney cells and native haptocorrin.
title_fullStr	Comparison of recombinant human haptocorrin expressed in human embryonic kidney cells and native haptocorrin.
title_full_unstemmed	Comparison of recombinant human haptocorrin expressed in human embryonic kidney cells and native haptocorrin.
title_sort	comparison of recombinant human haptocorrin expressed in human embryonic kidney cells and native haptocorrin.
publisher	Public Library of Science (PLoS)
publishDate	2012
url	https://doaj.org/article/e746c69532c449408e881ca48d381e85
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Comparison of recombinant human haptocorrin expressed in human embryonic kidney cells and native haptocorrin.

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