The collagen V homotrimer [alpha1(V)](3) production is unexpectedly favored over the heterotrimer [alpha1(V)](2)alpha2(V) in recombinant expression systems.

Journal of Biomedicine & Biotechnology
Muriel RouletF Ruggiero

Abstract

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 [alpha1(V)](2)alpha2(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 proalpha1(V) and proalpha2(V) chain cDNAs. Suprisingly, in all recombinant expression systems, the formation of [alpha1(V)](3) homotrimers was considerably favored over the heterotrimer. In addition, pepsin-sensitive proalpha2(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 alpha-chains showed that, contrary to fibroblasts, collagen V alpha-chains did not colocalized intracellularly in transfected cells. Monensin treatment had no effect on the heterotrimer pr...Continue Reading

References

Aug 15, 1992·Proceedings of the National Academy of Sciences of the United States of America·K VuoriK I Kivirikko
May 31, 1988·Biochemical and Biophysical Research Communications·K NagataK M Yamada
Jul 1, 1995·Matrix Biology : Journal of the International Society for Matrix Biology·A FichardF Ruggiero
Feb 3, 1997·Biochemical and Biophysical Research Communications·K FukudaY Yamada
Dec 31, 1997·The Journal of Biological Chemistry·A FichardF Ruggiero
Mar 21, 1998·The Journal of Biological Chemistry·S FrischholzK von der Mark
Mar 21, 1998·Matrix Biology : Journal of the International Society for Matrix Biology·M NokelainenK I Kivirikko
Apr 3, 1998·Matrix Biology : Journal of the International Society for Matrix Biology·S H McLaughlin, N J Bulleid
Dec 22, 1999·Seminars in Cell & Developmental Biology·S R Lamandé, J F Bateman
Jan 21, 2000·FEMS Microbiology Reviews·J L Cereghino, J M Cregg
Sep 28, 2000·Micron : the International Research and Review Journal for Microscopy·D E Birk
May 19, 2001·The Journal of Biological Chemistry·E KesslerF Ruggiero
Jun 26, 2001·The Journal of Biological Chemistry·H Chanut-DelalandeF Ruggiero
Nov 19, 2003·Advanced Drug Delivery Reviews·David OlsenJames Polarek
Mar 3, 2004·The Journal of Biological Chemistry·Simon StephanCay M Kielty
Jun 17, 2004·Molecular and Cellular Biology·Hélène Chanut-DelalandeFlorence Ruggiero
Sep 24, 2004·The Journal of Biological Chemistry·Richard J WenstrupDavid E Birk
Mar 8, 2005·Matrix Biology : Journal of the International Society for Matrix Biology·Antje NeubauerJohanna Myllyharju
Aug 9, 2005·Pathologie-biologie·Sylvie Ricard-Blum, Florence Ruggiero
Apr 5, 2007·The Biochemical Journal·Christelle Bonod-BidaudFlorence Ruggiero
Apr 30, 2008·Methods : a Companion to Methods in Enzymology·Florence Ruggiero, Manuel Koch

❮ Previous
Next ❯

Citations

Oct 13, 2012·The Journal of Biological Chemistry·Chenxi YangDaniel S Greenspan
Jan 21, 2012·Biomedical Materials·Jerome A Werkmeister, John A M Ramshaw
Jul 15, 2020·Experimental Eye Research·Edgar M Espana, David E Birk

❮ Previous
Next ❯

Methods Mentioned

BETA
PCR
transfection

Software Mentioned

Quantity One

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.