Insect Cell Glycosylation and Its Impact on the Functionality of a Recombinant Intracrystalline Nacre Protein, AP24

Biochemistry
Eric P ChangJohn Spencer Evans

Abstract

The impacts of glycosylation on biomineralization protein function are largely unknown. This is certainly true for the mollusk shell, where glycosylated intracrystalline proteins such as AP24 (Haliotis rufescens) exist but their functions and the role of glycosylation remain elusive. To assess the effect of glycosylation on protein function, we expressed two recombinant variants of AP24: an unglycosylated bacteria-expressed version (rAP24N) and a glycosylated insect cell-expressed version (rAP24G). Our findings indicate that rAP24G is expressed as a single polypeptide containing variations in glycosylation that create microheterogeneity in rAP24G molecular masses. These post-translational modifications incorporate O- and N-glycans and anionic monosialylated and bisialylated, and monosulfated and bisulfated monosaccharides on the protein molecules. AFM and DLS experiments confirm that both rAP24N and rAP24G aggregate to form protein phases, with rAP24N exhibiting a higher degree of aggregation, compared to rAP24G. With regard to functionality, we observe that both recombinant proteins exhibit similar behavior within in vitro calcium carbonate mineralization assays and potentiometric titrations. However, rAP24G modifies crystal g...Continue Reading

References

Aug 1, 1976·Proceedings of the National Academy of Sciences of the United States of America·S WeinerL Hood
May 1, 1980·The Journal of Experimental Medicine·T HaliotisR B Herberman
Dec 1, 1981·Molecular Immunology·R J Robb, K A Smith
Nov 1, 1981·Journal of Virological Methods·M TrudelP Payment
Apr 4, 2002·Proceedings of the National Academy of Sciences of the United States of America·Martina SchwarzkopfRüdiger Horstkorte
Nov 25, 2003·Journal of Microscopy·S BlankM Fritz
May 7, 2005·Nature Biotechnology·Thomas A KostDonald L Jarvis
Feb 28, 2007·BMC Bioinformatics·Oscar Conchillo-SoléSalvador Ventura
Mar 27, 2007·Biomacromolecules·Sebastiano Collino, John Spencer Evans
Sep 24, 2008·Chembiochem : a European Journal of Chemical Biology·Benjamin MarieFrédéric Marin
Oct 10, 2008·BMC Genomics·A Keith DunkerVladimir N Uversky
Nov 29, 2008·BMC Bioinformatics·Stephen E Hamby, Jonathan D Hirst
Mar 5, 2009·Physical Review Letters·Xiaodong Li, Zaiwang Huang
Aug 15, 2009·Science·Michio SuzukiHiromichi Nagasawa
Feb 6, 2010·Proceedings of the National Academy of Sciences of the United States of America·Lukasz GoldschmidtDavid Eisenberg
Feb 23, 2010·Micron : the International Research and Review Journal for Microscopy·Antonino NatoliWerner E G Müller
Mar 3, 2010·Nature Methods·Mikael Oliveberg
Aug 24, 2011·Physical Chemistry Chemical Physics : PCCP·Andreas VerchHelmut Cölfen
Oct 11, 2011·Journal of Structural Biology·Antonio G ChecaMarc-Georg Willinger
Oct 20, 2011·Molecular BioSystems·Franck PeysselonSylvie Ricard-Blum
Dec 22, 2011·Nature Communications·Raffaella DemichelisDenis Gebauer
Apr 5, 2012·Chembiochem : a European Journal of Chemical Biology·Paula Ramos-SilvaFrédéric Marin
Apr 28, 2012·Gene·Chihiro NogawaTetsuro Samata
May 26, 2012·BMC Bioinformatics·Lukasz P Kozlowski, Janusz M Bujnicki
Dec 6, 2012·Proceedings of the National Academy of Sciences of the United States of America·Benjamin MarieCaroline Montagnani
Jan 29, 2013·Journal of Oral and Maxillofacial Surgery : Official Journal of the American Association of Oral and Maxillofacial Surgeons·Louis Mandel
Apr 16, 2013·The EMBO Journal·Catharina SteentoftHenrik Clausen
Jul 11, 2013·PloS One·Jagat Singh ChauhanGajendra P S Raghava
Jul 20, 2013·Biochemistry·Iva PerovicJohn Spencer Evans
Jan 25, 2014·Chemical Society Reviews·Denis GebauerHelmut Cölfen

❮ Previous
Next ❯

Citations

May 17, 2017·Biophysics Reviews·Ashit Rao, Helmut Cölfen
Oct 17, 2019·Briefings in Bioinformatics·Akila KatuwawalaLukasz Kurgan
Jun 29, 2021·Comparative Biochemistry and Physiology. Part B, Biochemistry & Molecular Biology·Crisalejandra Rivera-Pérez, Norma Y Hernández-Saavedra

❮ Previous
Next ❯

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.