Direct site-selective covalent protein immobilization catalyzed by a phosphopantetheinyl transferase

Journal of the American Chemical Society
Lu Shin WongJason Micklefield

Abstract

Immobilization of proteins onto solid supports is important in the preparation of functional protein microarrays and in the development of bead-based bioassays, biosensors, and industrial biocatalysts. In order to generate the stable, functional, and homogeneous materials required for these applications, attention has focused on methods that enable the efficient and site-specific covalent immobilization of recombinant proteins onto a wide range of platforms. To this end, the phosphopantetheinyl transferase Sfp was employed to catalyze the direct immobilization of recombinant proteins bearing the small, genetically encoded ybbR tag onto surfaces functionalized with CoA. Using mass spectrometry, it was shown that the Sfp catalyzes immobilization of a model acyl carrier protein (ACP) onto CoA-derivatized PEGA resin beads through specific covalent bond formation. Luciferase (Luc) and glutathione-S-transferase (GST) ybbR-fusion proteins were similarly immobilized onto PEGA resin retaining high levels of enzyme activity. This strategy was also successfully applied for the immobilization of the ACP, as well as ybbR-Luc, -GST, and -thioredoxin fusion proteins, on hydrogel microarray slides. Overall, the Sfp-catalyzed surface ligation i...Continue Reading

Citations

Jan 21, 2009·Analytical and Bioanalytical Chemistry·Changjiang YouJacob Piehler
Jul 24, 2012·Bioconjugate Chemistry·Kosuke MinamihataNoriho Kamiya
Jul 11, 2013·Bioconjugate Chemistry·Mohammad RashidianMark D Distefano
Mar 1, 2012·Journal of the American Chemical Society·Nathan D ShapiroGeorge M Whitesides
May 16, 2009·Journal of the American Chemical Society·Yekui Zou, Jun Yin
Apr 9, 2010·Journal of the American Chemical Society·Katarzyna A MosiewiczMatthias P Lutolf
Sep 18, 2012·Nature Methods·Nicolas M KosaMichael D Burkart
Jan 13, 2011·Organic & Biomolecular Chemistry·Xuxu YanKarine Auclair
Oct 20, 2010·Molecular & Cellular Proteomics : MCP·Christian M LochJames E Strickler
Jul 26, 2012·Biotechnology Journal·Takuya MatsumotoAkihiko Kondo
Dec 22, 2016·Journal of the American Chemical Society·Kanwal S PallaMatthew B Francis
Aug 21, 2015·Chemical Communications : Chem Comm·Yun-Kun QiJi-Shen Zheng
Mar 16, 2018·Chemical Communications : Chem Comm·Xiafeng WangZhengying Pan
Feb 28, 2018·Biotechnology Journal·Jaehyun Cha, Inchan Kwon
Jan 12, 2013·International Journal of Molecular Sciences·Raushan Kumar SinghJung-Kul Lee
May 31, 2016·Angewandte Chemie·Christian B RosenMatthew B Francis
Aug 14, 2009·Organic & Biomolecular Chemistry·Murat Sunbul, Jun Yin
Sep 10, 2009·Chembiochem : a European Journal of Chemical Biology·Keitaro EbisuMasumi Taki
Jan 14, 2010·Angewandte Chemie·Dirk WeinrichHerbert Waldmann
Sep 15, 2010·Chemistry : a European Journal·Lu Shin WongJason Micklefield
Apr 24, 2009·Expert Review of Proteomics·Oda StoevesandtMingyue He
Sep 25, 2012·Chemical Communications : Chem Comm·Long YiHerbert Waldmann
Sep 12, 2015·Physical Biology·Kamila KlameckaHeinrich Leonhardt
Nov 28, 2012·Chembiochem : a European Journal of Chemical Biology·Anna-Winona StruckJason Micklefield
Feb 6, 2010·Organic & Biomolecular Chemistry·Lu Shin WongJason Micklefield
Mar 7, 2014·Chembiochem : a European Journal of Chemical Biology·Hsiao-Chan WangChun-Cheng Lin
Oct 16, 2015·Chembiochem : a European Journal of Chemical Biology·Fumihiro IshikawaHideaki Kakeya
Aug 16, 2013·Chemical Communications : Chem Comm·Dennis M BauerLjiljana Fruk
Apr 8, 2010·Langmuir : the ACS Journal of Surfaces and Colloids·Sinem EnginDoris Wedlich
Jul 4, 2009·Chemical Reviews·Lu Shin WongJason Micklefield
Jan 23, 2010·Analytical Chemistry·Sharon WaichmanJacob Piehler
Jun 5, 2014·ACS Nano·Diana A PippigHermann E Gaub
Oct 24, 2014·Bioconjugate Chemistry·Yiqing ZhouZhengying Pan

❮ 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.