The Smallest Isoform of Metridia longa Luciferase as a Fusion Partner for Hybrid Proteins.

International Journal of Molecular Sciences
Marina D LarionovaEugene S Vysotski

Abstract

Bioluminescent proteins are widely used as reporter molecules in various in vitro and in vivo assays. The smallest isoform of Metridia luciferase (MLuc7) is a highly active, naturally secreted enzyme which, along with other luciferase isoforms, is responsible for the bright bioluminescence of marine copepod Metridia longa. In this study, we report the construction of two variants of a hybrid protein consisting of MLuc7 and 14D5a single-chain antibody to the surface glycoprotein E of tick-borne encephalitis virus as a model fusion partner. We demonstrate that, whereas fusion of a single-chain antibody to either N- or C-terminus of MLuc7 does not affect its bioluminescence properties, the binding site on the single-chain antibody influences its binding capacity. The affinity of 14D5a-MLuc7 hybrid protein (KD = 36.2 nM) where the C-terminus of the single-chain antibody was fused to the N-terminus of MLuc7, appeared to be 2.5-fold higher than that of the reverse, MLuc7-14D5a (KD = 87.6 nM). The detection limit of 14D5a-MLuc7 hybrid protein was estimated to be 45 pg of the recombinant glycoprotein E. Although the smallest isoform of M. longa luciferase was tested as a fusion partner only with a single-chain antibody, it is reasonabl...Continue Reading

References

Jan 11, 1977·Biochemistry·J C MatthewsM J Cormier
Jul 17, 1975·Nature·O Shimomura, F H Johnson
Feb 28, 2001·Fresenius' Journal of Analytical Chemistry·J C Lewis, S Daunert
Jan 18, 2003·Cellular and Molecular Life Sciences : CMLS·V R Viviani
Jan 6, 2005·The Journal of Biological Chemistry·Yves J M BollenCarlo P M van Mierlo
Jul 22, 2006·Protein Engineering, Design & Selection : PEDS·Andreas Markus LoeningSanjiv Sam Gambhir
Jun 20, 2007·Molecular Imaging and Biology : MIB : the Official Publication of the Academy of Molecular Imaging·Katy M VenisnikAnna M Wu
Nov 6, 2007·Biochemical and Biophysical Research Communications·Satoshi Inouye, Yuiko Sahara
Jan 25, 2008·BMC Bioinformatics·Yang Zhang
Feb 12, 2008·Photochemical & Photobiological Sciences : Official Journal of the European Photochemistry Association and the European Society for Photobiology·Maxim S TitushinEugene S Vysotski
Aug 30, 2008·Photochemical & Photobiological Sciences : Official Journal of the European Photochemistry Association and the European Society for Photobiology·Vasilisa V BorisovaEugene S Vysotski
Oct 27, 2009·Biochemical and Biophysical Research Communications·Kedar G PatelJames R Swartz
Sep 17, 2011·Biotechnology Advances·Bakhos A Tannous, Jian Teng
Dec 6, 2011·Biochemical and Biophysical Research Communications·Svetlana V MarkovaEugene S Vysotski
Mar 26, 2013·Photochemical & Photobiological Sciences : Official Journal of the European Photochemistry Association and the European Society for Photobiology·Elena V EremeevaEugene S Vysotski
Jan 28, 2014·Journal of Immunological Methods·Julia UstinovaRaivo Uibo
Dec 30, 2014·Biochemical and Biophysical Research Communications·Svetlana V MarkovaEugene S Vysotski
Dec 31, 2014·Nature Methods·Jianyi YangYang Zhang
May 1, 2015·Analytical and Bioanalytical Chemistry·Ludmila P BurakovaLudmila A Frank
Nov 5, 2015·Biochemistry. Biokhimii︠a︡·S V Markova, E S Vysotski
Nov 1, 2016·Photochemistry and Photobiology·Daria V Smirnova, Natalia N Ugarova
Dec 15, 2016·Biochemical and Biophysical Research Communications·Marina D LarionovaEugene S Vysotski
Dec 16, 2016·Photochemistry and Photobiology·Marina D LarionovaEugene S Vysotski
Aug 29, 2017·Journal of Photochemistry and Photobiology. B, Biology·Svetlana V MarkovaEugene S Vysotski
May 14, 2018·Journal of Photochemistry and Photobiology. B, Biology·Marina D LarionovaEugene S Vysotski
Dec 27, 2018·Photochemistry and Photobiology·Svetlana V MarkovaEugene S Vysotski
Feb 15, 2019·Scientific Reports·Ramakrishnan GopalakrishnanPreet M Chaudhary
Feb 20, 2019·Nature Biotechnology·José Juan Almagro ArmenterosHenrik Nielsen

❮ Previous
Next ❯

Datasets Mentioned

BETA
MT683623
MT683622

Methods Mentioned

BETA
ELISA
affinity purification
PCR
genetic modification
transfection
protein assay

Software Mentioned

SignalP

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.