Abstract
Immobilization of functional proteins such as enzymes on solid surfaces produces a variety of effects ranging from the reversal and strong inhibition to the enhancement of protein stability and function. Such effects are protein-dependent and are affected by the physical and chemical properties of the surfaces. Functional consequences of protein immobilization on the surface of gold nanoparticles (AuNPs) are protein-dependent and require thorough investigation using suitable functional tests. However, traditional approaches to making control samples, i.e., immobilized protein vs. protein in solution in absence of any nanoparticles do not provide sufficiently identical reaction conditions and complicate interpretation of the results. This report provides advice and methods for preparing AuNP-conjugated preparations generally suitable for studying the effects of immobilization on the activity and stability of different functional proteins. We use bovine catalase to illustrate our approach, but the methods are easily adaptable to any other enzyme or protein. The AuNP-immobilized enzyme showed increased stability at elevated temperatures compared to the same enzyme in solution.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Faulk WP, Taylor GM (1971) An immunocolloid method for the electron microscope. Immunochemistry 8(11):1081–1083
Neagu C, van der Werf KO, Putman CAJ et al (1994) Analysis of immunolabeled cells by atomic force microscopy, optical microscopy, and flow cytometry. J Struct Biol 112(1):32–40
Zhao W, Chiuman W, Brook MA et al (2007) Simple and rapid colorimetric biosensors based on DNA aptamer and noncrosslinking gold nanoparticle aggregation. Chembiochem 8(7):727–731
Medley CD, Smith JE, Tang Z et al (2008) Gold nanoparticle-based colorimetric assay for the direct detection of cancerous cells. Anal Chem 80(4):1067–1072
Reynolds RA III, Mirkin CA, Letsinger RL (2000) A gold nanoparticle/latex microsphere-based colorimetric oligonucleotide detection method. Pure Appl Chem 72(1–2):229–235
Choi DH, Lee SK, Oh YK et al (2010) A dual gold nanoparticle conjugate-based lateral flow assay (LFA) method for the analysis of troponin I. Biosens Bioelectron 25(8):1999–2002
Rong-Hwa S, Shiao-Shek T, Der-Jiang C et al (2010) Gold nanoparticle-based lateral flow assay for detection of staphylococcal enterotoxin B. Food Chem 188(2):462–466
Girotti S, Eremin S, Montoya A et al (2010) Development of a chemiluminescent ELISA and a colloidal gold-based LFIA for TNT detection. Anal Bioanal Chem 396:687–695
Yu CY, Ang GY, Chua AL et al (2011) Dry-reagent gold nanoparticle-based lateral flow biosensor for the simultaneous detection of Vibrio cholerae serogroups O1 and O139. J Microbiol Methods 86(3):277–282
Schultz DA (2003) Plasmon resonant particles for biological detection. Curr Opin Biotechnol 14(1):13–22
Tanaka M, Matsuo K, Enomoto M et al (2004) A sol particle homogeneous immunoassay for measuring serum cystatin C. Clin Biochem 37(1):27–35
Thanh NTK, Rosenzweig Z (2002) Development of an aggregation-based immunoassay for anti-protein A using gold nanoparticles. Anal Chem 74(7):1624–1628
Thanh NTK, Rees JH, Rosenzweig Z (2002) Laser-based double beam absorption detection for aggregation immunoassays using gold nanoparticles. Anal Bioanal Chem 374(7–8):1174–1178
Zhang CX, Zhang Y, Wang X et al (2003) Hyper-Rayleigh scattering of protein-modified gold nanoparticles. Anal Biochem 320(1):136–140
Otsuka H, Akiyama Y, Nagasaki Y et al (2002) Quantitative and reversible lectin-induced association of gold nanoparticles modified with alpha-lactosyl-omega-mercapto-poly(ethylene glycol). J Am Chem Soc 123(34):8226–8230
Torrecilla JS, Mena LM, Yanez SP et al (2008) A neural network approach based on gold-nanoparticle enzyme biosensor. J Chemometr 22(1):46–53
Pingarro JM, Yanez SP, Gonzalez-Cortes A (2008) Gold nanoparticle-based electrochemical biosensors. Electrochim Acta 53(19):5848–5866
Li J, Song S, Liu X et al (2008) Enzyme-based multi-component optical nanoprobes for sequence-specific detection of DNA hybridization. Adv Mater 20(3):497–500
Giulio FP, Kingston DGI, Tamarkin L (2006) Colloidal gold nanoparticles: A novel nanoparticle platform for developing multifunctional tumor-targeted drug delivery vectors. Drug Devel Res 67(1):47–54
Jun HK, Lee TR (2006) Discrete thermally responsive hydrogel-coated gold nanoparticles for use as drug-delivery vehicles. Drug Devel Res 67(1):61–69
Connolly S, Fitzmaurice D (1999) Programmed assembly of gold nanocrystals in aqueous solution. Adv Mater 11(14):1202–1205
Park SJ, Lazarides AA, Mirkin CA et al (2001) Directed assembly of periodic materials from protein and oligonucleotide-modified nanoparticle building blocks. Angew Chem Int Ed Engl 40(15):2909–2912
Niemeyer CM (2001) Nanoparticles, proteins, and nucleic acids: biotechnology meets materials science. Angew Chem Int Ed Engl 40(22):4128–4158
Mastroianni AJ, Claridge SA, Alivisatos AP (2009) Pyramidal and chiral groupings of gold nanocrystals assembled using DNA scaffolds. J Am Chem Soc 131(24):8455–8459
Ferrari E, Darios F, Zhang F et al (2010) Binary polypeptide system for permanent and oriented protein immobilization. J Nanobiotechnology 8:9
Brust M, Walker M, Bethell D et al (1994) Synthesis of thiol-derivatised gold nanoparticles in a two-phase liquid–liquid system. J Chem Soc Chem Commun 7(7):801–802
Perrault SD, Chan WC (2009) Synthesis and surface modification of highly monodispersed, spherical gold nanoparticles of 50-200 nm. J Am Chem Soc 131(47):17042–17043
Martin MN, Basham JI, Chando P et al (2010) Charged gold nanoparticles in non-polar solvents: 10-min synthesis and 2D self-assembly. Langmuir 26(10):7410–7417
Zhao W, Lin L, Hsing I (2009) Rapid synthesis of DNA-functionalized gold nanoparticles in salt solution using mononucleotide-Âmediated conjugation. Biconjug Chem 20(6):1218–1222
Wangoo N, Bhasin KK, Mehta SK et al (2008) Synthesis and capping of water-dispersed gold nanoparticles by an amino acid: bioconjugation and binding studies. J Colloid Interface Sci 323(2):247–254
Satyavani K, Gurudeeban S, Ramanathan T et al (2011) Biomedical potential of silver nanoparticles synthesized from calli cells of Citrullus colocynthis (L.) Schrad. J Nanobiotechnol 9:43
Walter JG, Petersen S, Stahl F et al (2010) Laser ablation-based one-step generation and bio-functionalization of gold nanoparticles conjugated with aptamers. J Nanobiotechnol 8:21
Conde J, de la Fuente JM, Baptista PV (2010) RNA quantification using gold nanoprobes—application to cancer diagnostics. J Nanobiotechnol 8:5
Crumbliss AL, Stonehuerner J, Henkens RW et al (1994) The use of inorganic materials to control or maintain immobilized enzyme activity. New J Chem 18:327–339
Gole A, Dash C, Ramakrishnan V et al (2001) Pepsin–gold colloid conjugates: preparation, characterization, and enzymatic activity. Langmuir 17(5):1674–1679
Huang F, Huang CC, Chang HT (2003) Exploring the activity and specificity of gold nanoparticle-bound trypsin by capillary electrophoresis with laser-induced fluorescence detection. Langmuir 19(18):7498–7502
Lv M, Zhu E, Su Y et al (2009) Trypsin-gold nanoparticle conjugates: binding, enzymatic activity, and stability. Prep Biochem Biotechnol 39(4):429–438
Li H, Huang J, Lv J et al (2005) Nanoparticle PCR: Nanogold-assisted PCR with enhanced specificity. Angew Chem Int Ed Engl 44(32):5100–5103
Li M, Lin YC, Wu CC et al (2005) Enhancing the efficiency of a PCR using gold nanoparticles. Nucl Acids Res 33(21):184
Asuri P, Karajanagi SS, Yang H et al (2006) Increasing protein stability through control of the nanoscale environment. Langmuir 22(13):5833–5836
Halling PJ, Ulijn RV, Flitsch SL (2005) Understanding enzyme action on immobilised substrates. Curr Opin Biotechnol 16(4):385–392
Basso A, Braiuca P, Ebert C et al (2006) Properties and applications of supports for enzyme-mediated transformations in solid phase synthesis. J Chem Technol Biotechnol 81(10):1626–1640
Cortez J, Vorobieva E, Gralheira D et al (2011) Bionanoconjugates of tyrosinase and peptide-derivatised gold nanoparticles for biosensing of phenolic compounds. J Nanopart Res 13:1101–1113
Roach P, Farrar D, Perry CC (2006) Surface tailoring for controlled protein adsorption: effect of topography at the nanometer scale and chemistry. J Am Chem Soc 128(12):3939–3945
Hong R, Fischer NO, Verma A et al (2004) Control of protein structure and function through surface recognition by tailored nanoparticle scaffolds. J Am Chem Soc 126(3):739–743
Karajanagi SS, Vertegel AA, Kane RS et al (2004) Structure and function of enzymes adsorbed onto single-walled carbon nanotubes. Langmuir 20(26):11594–11599
Gole A, Dash C, Soman C et al (2001) On the preparation, characterization, and enzymatic activity of fungal protease-gold colloid bioconjugates. Bioconjug Chem 12(5):684–690
Keating CD, Kovaleski KM, Natan MJJ (1998) Protein:colloid conjugates for surface-enhanced Raman scattering: stability and control of Âprotein orientation. Phys Chem B 102(47):9404–9413
Dykman LA, Bogatyrev VA, Khlebtsov BN et al (2005) A protein assay based on colloidal gold conjugates with trypsin. Anal Biochem 341(1):16–21
Laera S, Ceccone G, Rossi F et al (2011) Measuring protein structure and stability of protein–nanoparticle systems with synchrotron radiation circular dichroism. Nano Lett 11(10):4480–4484
Aubin-Tam ME, Hamad-Schifferli K (2005) Gold nanoparticle  −  cytochrome C complexes: the effect of nanoparticle ligand charge on protein structure. Langmuir 21(26):12080–12084
Kolobanova SV, Filippova IY, Lysogorskaya EN (2001) The enzymatic segment condensation of peptides on a solid phase in organic medium. Bioorg Khim 27(5):347–351
Doeze RHP, Maltman BA, Egan CL et al (2004) Profiling primary protease specificity by peptide synthesis on a solid support. Angew Chem Int Ed Engl 43(24):3138–3141
Zamocky M, Koller F (1999) Understanding the structure and function of catalases: clues from molecular evolution and in vitro mutagenesis. Prog Biophys Mol Biol 72(1):19–66
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2012 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Bailes, J., Gazi, S., Ivanova, R., Soloviev, M. (2012). Effect of Gold Nanoparticle Conjugation on the Activity and Stability of Functional Proteins. In: Soloviev, M. (eds) Nanoparticles in Biology and Medicine. Methods in Molecular Biology, vol 906. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-953-2_7
Download citation
DOI: https://doi.org/10.1007/978-1-61779-953-2_7
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-61779-952-5
Online ISBN: 978-1-61779-953-2
eBook Packages: Springer Protocols