Glutathione-S-transferase Fusion Protein Nanosensor

Overview

Journal Nano Lett

Publisher American Chemical Society

Specialty Biotechnology

Date 2020 Sep 21

PMID 32955895

Citations 4

Authors

Ryan M Williams

Jackson D Harvey

Januka Budhathoki-Uprety

Daniel A Heller

Affiliations

Soon will be listed here.

Abstract

Fusion protein tags are widely used to capture and track proteins in research and industrial bioreactor processes. Quantifying fusion-tagged proteins normally requires several purification steps coupled with classical protein assays. Here, we developed a broadly applicable nanosensor platform that quantifies glutathione-S-transferase (GST) fusion proteins in real-time. We synthesized a glutathione-DNA-carbon nanotube system to investigate glutathione-GST interactions via semiconducting single-walled carbon nanotube (SWCNT) photoluminescence. We found that SWCNT fluorescence wavelength and intensity modulation occurred specifically in response to GST and GST-fusions. The sensor response was dependent on SWCNT structure, wherein ( - , 3) = 1 nanotube wavelength and intensity responses correlated with nanotube diameter distinctly from ( - , 3) = 2 SWCNT responses. We also found broad functionality of this sensor to diverse GST-tagged proteins. This work comprises the first label-free optical sensor for GST and has implications for the assessment of protein expression in situ, including in imaging and industrial bioreactor settings.

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References

Moore R, Hill E, Horan T, Yano N, Kim K, MacLaughlan S . HE4 (WFDC2) gene overexpression promotes ovarian tumor growth. Sci Rep. 2014; 4:3574. PMC: 3880958. DOI: 10.1038/srep03574. View

Lichty J, Malecki J, Agnew H, Michelson-Horowitz D, Tan S . Comparison of affinity tags for protein purification. Protein Expr Purif. 2005; 41(1):98-105. DOI: 10.1016/j.pep.2005.01.019. View

Goldring J . Protein quantification methods to determine protein concentration prior to electrophoresis. Methods Mol Biol. 2012; 869:29-35. DOI: 10.1007/978-1-61779-821-4_3. View

Yang R, Tang Z, Yan J, Kang H, Kim Y, Zhu Z . Noncovalent assembly of carbon nanotubes and single-stranded DNA: an effective sensing platform for probing biomolecular interactions. Anal Chem. 2008; 80(19):7408-13. DOI: 10.1021/ac801118p. View

Iverson N, Barone P, Shandell M, Trudel L, Sen S, Sen F . In vivo biosensing via tissue-localizable near-infrared-fluorescent single-walled carbon nanotubes. Nat Nanotechnol. 2013; 8(11):873-80. PMC: 4066962. DOI: 10.1038/nnano.2013.222. View

Kharas M, Lengner C, Al-Shahrour F, Bullinger L, Ball B, Zaidi S . Musashi-2 regulates normal hematopoiesis and promotes aggressive myeloid leukemia. Nat Med. 2010; 16(8):903-8. PMC: 3090658. DOI: 10.1038/nm.2187. View

OConnell M, Bachilo S, Huffman C, Moore V, Strano M, Haroz E . Band gap fluorescence from individual single-walled carbon nanotubes. Science. 2002; 297(5581):593-6. DOI: 10.1126/science.1072631. View

Heller D, Pratt G, Zhang J, Nair N, Hansborough A, Boghossian A . Peptide secondary structure modulates single-walled carbon nanotube fluorescence as a chaperone sensor for nitroaromatics. Proc Natl Acad Sci U S A. 2011; 108(21):8544-9. PMC: 3102399. DOI: 10.1073/pnas.1005512108. View

Bell M, Engleka M, Malik A, Strickler J . To fuse or not to fuse: what is your purpose?. Protein Sci. 2013; 22(11):1466-77. PMC: 3831663. DOI: 10.1002/pro.2356. View

10.

So H, Won K, Kim Y, Kim B, Ryu B, Na P . Single-walled carbon nanotube biosensors using aptamers as molecular recognition elements. J Am Chem Soc. 2005; 127(34):11906-7. DOI: 10.1021/ja053094r. View

11.

Smith D, Corcoran L . Expression and purification of glutathione-S-transferase fusion proteins. Curr Protoc Mol Biol. 2008; Chapter 16:Unit16.7. DOI: 10.1002/0471142727.mb1607s28. View

12.

Huang J, Li J, Lyu Y, Miao Q, Pu K . Molecular optical imaging probes for early diagnosis of drug-induced acute kidney injury. Nat Mater. 2019; 18(10):1133-1143. DOI: 10.1038/s41563-019-0378-4. View

13.

Bisker G, Bakh N, Lee M, Ahn J, Park M, OConnell E . Insulin Detection Using a Corona Phase Molecular Recognition Site on Single-Walled Carbon Nanotubes. ACS Sens. 2018; 3(2):367-377. DOI: 10.1021/acssensors.7b00788. View

14.

Roxbury D, Jena P, Shamay Y, Horoszko C, Heller D . Cell Membrane Proteins Modulate the Carbon Nanotube Optical Bandgap via Surface Charge Accumulation. ACS Nano. 2015; 10(1):499-506. PMC: 4975035. DOI: 10.1021/acsnano.5b05438. View

15.

Yi H, Ghosh D, Ham M, Qi J, Barone P, Strano M . M13 phage-functionalized single-walled carbon nanotubes as nanoprobes for second near-infrared window fluorescence imaging of targeted tumors. Nano Lett. 2012; 12(3):1176-1183. PMC: 3905603. DOI: 10.1021/nl2031663. View

16.

Cheng P, Miao Q, Li J, Huang J, Xie C, Pu K . Unimolecular Chemo-fluoro-luminescent Reporter for Crosstalk-Free Duplex Imaging of Hepatotoxicity. J Am Chem Soc. 2019; 141(27):10581-10584. DOI: 10.1021/jacs.9b02580. View

17.

Bisker G, Dong J, Park H, Iverson N, Ahn J, Nelson J . Protein-targeted corona phase molecular recognition. Nat Commun. 2016; 7:10241. PMC: 4729864. DOI: 10.1038/ncomms10241. View

18.

Barman U, Mukhopadhyay G, Goswami N, Ghosh S, Paily R . Detection of Glutathione by Glutathione-S-Transferase-Nanoconjugate Ensemble Electrochemical Device. IEEE Trans Nanobioscience. 2017; 16(4):271-279. DOI: 10.1109/TNB.2017.2698241. View

19.

Kim M, Wu X, Ao G, He X, Kwon H, Hartmann N . Mapping Structure-Property Relationships of Organic Color Centers. Chem. 2019; 4(9):2180-2191. PMC: 6874404. DOI: 10.1016/j.chempr.2018.06.013. View

20.

Boyer T, Vessey D, Holcomb C, Saley N . Studies of the relationship between the catalytic activity and binding of non-substrate ligands by the glutathione S-transferases. Biochem J. 1984; 217(1):179-85. PMC: 1153195. DOI: 10.1042/bj2170179. View