Improved secretion of glycoproteins using an N-glycan-restricted passport sequence tag recognized by cargo receptor
Open Access
- 13 March 2020
- journal article
- research article
- Published by Springer Science and Business Media LLC in Nature Communications
- Vol. 11 (1), 1-9
- https://doi.org/10.1038/s41467-020-15192-1
Abstract
MCFD2 and ERGIC-53, which are the products of causative genes of combined factor V and factor VIII deficiency, form a cargo receptor complex responsible for intracellular transport of these coagulation factors in the early secretory pathway. In this study, using an NMR technique, we successfully identified an MCFD2-binding segment from factor VIII composed of a 10 amino acid sequence that enhances its secretion. This prompted us to examine possible effects of attaching this sequence to recombinant glycoproteins on their secretion. We found that the secretion level of recombinant erythropoietin was significantly increased simply by tagging it with the passport sequence. Our findings not only provide molecular basis for the intracellular trafficking of coagulation factors and their genetic deficiency but also offer a potentially useful tool for increasing the production yields of recombinant glycoproteins of biopharmaceutical interest.Funding Information
- MEXT | Japan Society for the Promotion of Science (JP17H06414, JP25102008, JP24249002)
This publication has 31 references indexed in Scilit:
- Fiji: an open-source platform for biological-image analysisNature Methods, 2012
- Synchronization of secretory protein traffic in populations of cellsNature Methods, 2012
- HNK-1 Epitope-carrying Tenascin-C Spliced Variant Regulates the Proliferation of Mouse Embryonic Neural Stem Cells*Online Journal of Public Health Informatics, 2010
- Structural basis for the cooperative interplay between the two causative gene products of combined factor V and factor VIII deficiencyProceedings of the National Academy of Sciences of the United States of America, 2010
- Functional roles of the factor VIII B domainHaemophilia, 2009
- Recent developments in the understanding of the combined deficiency of FV and FVIIIBritish Journal of Haematology, 2009
- Genotype-phenotype correlation in combined deficiency of factor V and factor VIIIBlood, 2008
- The ER-Golgi intermediate compartment (ERGIC): in search of its identity and functionJournal of Cell Science, 2006
- Bleeding due to disruption of a cargo-specific ER-to-Golgi transport complexNature Genetics, 2003
- Identification, by a monoclonal antibody, of a 53-kD protein associated with a tubulo-vesicular compartment at the cis-side of the Golgi apparatus.The Journal of cell biology, 1988