The Collagen V Homotrimer [α1(V)]3 Production Is Unexpectedly Favored over the Heterotrimer [α1(V)]2α2(V) in Recombinant Expression Systems
Joint Authors
Bonod-Bidaud, Christelle
Chanut-Delalande, Hélène
Välkkilä, Merja
Ala-Kokko, Leena
Hämäläinen, Eija-Riitta
Kessler, Efrat
Männikkö, Minna
Ruggiero, Florence
Roulet, Muriel
Source
Journal of Biomedicine and Biotechnology
Issue
Vol. 2010, Issue 2010 (31 Dec. 2010), pp.1-13, 13 p.
Publisher
Hindawi Publishing Corporation
Publication Date
2010-06-27
Country of Publication
Egypt
No. of Pages
13
Main Subjects
Abstract EN
Collagen V, a fibrillar collagen with important functions in tissues, assembles into distinct chain associations.
The most abundant and ubiquitous molecular form is the heterotrimer [α1(V)]2α2(V).
In the attempt to produce high levels of recombinant collagen V heterotrimer for biomedical device uses, and to identify key factors that drive heterotrimeric chain association, several cell expression systems (yeast, insect, and mammalian cells) have been assayed by cotransfecting the human proα1(V) and proα2(V) chain cDNAs.
Suprisingly, in all recombinant expression systems, the formation of [α1(V)]3 homotrimers was considerably favored over the heterotrimer.
In addition, pepsin-sensitive proα2(V) chains were found in HEK-293 cell media indicating that these cells lack quality control proteins preventing collagen monomer secretion.
Additional transfection with Hsp47 cDNA, encoding the collagen-specific chaperone Hsp47, did not increase heterotrimer production.
Double immunofluorescence with antibodies against collagen V α-chains showed that, contrary to fibroblasts, collagen V α-chains did not colocalized intracellularly in transfected cells.
Monensin treatment had no effect on the heterotrimer production.
The heterotrimer production seems to require specific machinery proteins, which are not endogenously expressed in the expression systems.
The different constructs and transfected cells we have generated represent useful tools to further investigate the mechanisms of collagen trimer assembly.
American Psychological Association (APA)
Roulet, Muriel& Välkkilä, Merja& Chanut-Delalande, Hélène& Hämäläinen, Eija-Riitta& Kessler, Efrat& Ala-Kokko, Leena…[et al.]. 2010. The Collagen V Homotrimer [α1(V)]3 Production Is Unexpectedly Favored over the Heterotrimer [α1(V)]2α2(V) in Recombinant Expression Systems. Journal of Biomedicine and Biotechnology،Vol. 2010, no. 2010, pp.1-13.
https://search.emarefa.net/detail/BIM-467273
Modern Language Association (MLA)
Roulet, Muriel…[et al.]. The Collagen V Homotrimer [α1(V)]3 Production Is Unexpectedly Favored over the Heterotrimer [α1(V)]2α2(V) in Recombinant Expression Systems. Journal of Biomedicine and Biotechnology No. 2010 (2010), pp.1-13.
https://search.emarefa.net/detail/BIM-467273
American Medical Association (AMA)
Roulet, Muriel& Välkkilä, Merja& Chanut-Delalande, Hélène& Hämäläinen, Eija-Riitta& Kessler, Efrat& Ala-Kokko, Leena…[et al.]. The Collagen V Homotrimer [α1(V)]3 Production Is Unexpectedly Favored over the Heterotrimer [α1(V)]2α2(V) in Recombinant Expression Systems. Journal of Biomedicine and Biotechnology. 2010. Vol. 2010, no. 2010, pp.1-13.
https://search.emarefa.net/detail/BIM-467273
Data Type
Journal Articles
Language
English
Notes
Includes bibliographical references
Record ID
BIM-467273