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Title: Engineered (Lys) 6 -Tagged Recombinant Sulfide-Reactive Hemoglobin I for Covalent Immobilization at Multiwalled Carbon Nanotubes

Abstract

The recombinant HbI was fused with a poly-Lys tag ((Lys)6-tagged rHbI) for specific-site covalent immobilization on two carbon nanotube transducer surfaces, i.e., powder and vertically aligned carbon nanotubes. The immobilization was achieved by following two steps: (1) generation of amine-reactive ester from the carboxylic acid groups of the surfaces and (2) coupling these groups with the amine groups of the Lys-tag. We analyzed the immobilization process using different conditions and techniques to differentiate protein covalent attachment from physical adsorption. Fourier transform infrared microspectroscopy data showed a 14 cm–1 displacement of the protein’s amide I and amide II peaks to lower the frequency after immobilization. This result indicates a covalent attachment of the protein to the surface. Differences in the morphology of the carbon substrate with and without (Lys)6-tagged rHbI confirmed protein immobilization, as observed by transmission electron microscopy. The electrochemical studies, which were performed to evaluate the redox center of the immobilized protein, show a confinement suitable for an efficient electron transfer system. More importantly, the electrochemical studies allowed determination of a redox potential for the new (Lys)6-tagged rHbI. The data show that the protein is electrochemically active and retains its biological activity toward H2S.

Authors:
ORCiD logo [1];  [1];  [2];  [3];  [1]
+ Show Author Affiliations
  1. Department of Chemistry, P.O. Box 9000, University of Puerto Rico—Mayagüez Campus, Mayaguez 00680-9000, Puerto Rico
  2. Department of Chemistry, P.O. Box 372230, University of Puerto Rico—Cayey Campus, Cayey 00737-2230, Puerto Rico
  3. Department of Chemistry, P.O. Box 23346, University of Puerto Rico—Río Piedras Campus, San Juan 00931-3346, Puerto Rico
Publication Date:
Research Org.:
Brookhaven National Lab. (BNL), Upton, NY (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Basic Energy Sciences (BES)
OSTI Identifier:
1413569
Alternate Identifier(s):
OSTI ID: 1508298
Grant/Contract Number:  
SC0012704
Resource Type:
Published Article
Journal Name:
ACS Omega
Additional Journal Information:
Journal Name: ACS Omega Journal Volume: 2 Journal Issue: 12; Journal ID: ISSN 2470-1343
Publisher:
American Chemical Society
Country of Publication:
United States
Language:
English
Subject:
37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY

Citation Formats

Díaz-Ayala, Ramonita, Torres-González, Lisa, Pietri, Ruth, Cabrera, Carlos R., and López-Garriga, Juan. Engineered (Lys) 6 -Tagged Recombinant Sulfide-Reactive Hemoglobin I for Covalent Immobilization at Multiwalled Carbon Nanotubes. United States: N. p., 2017. Web. doi:10.1021/acsomega.7b01500.
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Díaz-Ayala, Ramonita, Torres-González, Lisa, Pietri, Ruth, Cabrera, Carlos R., & López-Garriga, Juan. Engineered (Lys) 6 -Tagged Recombinant Sulfide-Reactive Hemoglobin I for Covalent Immobilization at Multiwalled Carbon Nanotubes. United States. https://doi.org/10.1021/acsomega.7b01500
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Díaz-Ayala, Ramonita, Torres-González, Lisa, Pietri, Ruth, Cabrera, Carlos R., and López-Garriga, Juan. Fri . "Engineered (Lys) 6 -Tagged Recombinant Sulfide-Reactive Hemoglobin I for Covalent Immobilization at Multiwalled Carbon Nanotubes". United States. https://doi.org/10.1021/acsomega.7b01500.
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@article{osti_1413569,
title = {Engineered (Lys) 6 -Tagged Recombinant Sulfide-Reactive Hemoglobin I for Covalent Immobilization at Multiwalled Carbon Nanotubes},
author = {Díaz-Ayala, Ramonita and Torres-González, Lisa and Pietri, Ruth and Cabrera, Carlos R. and López-Garriga, Juan},
abstractNote = {The recombinant HbI was fused with a poly-Lys tag ((Lys)6-tagged rHbI) for specific-site covalent immobilization on two carbon nanotube transducer surfaces, i.e., powder and vertically aligned carbon nanotubes. The immobilization was achieved by following two steps: (1) generation of amine-reactive ester from the carboxylic acid groups of the surfaces and (2) coupling these groups with the amine groups of the Lys-tag. We analyzed the immobilization process using different conditions and techniques to differentiate protein covalent attachment from physical adsorption. Fourier transform infrared microspectroscopy data showed a 14 cm–1 displacement of the protein’s amide I and amide II peaks to lower the frequency after immobilization. This result indicates a covalent attachment of the protein to the surface. Differences in the morphology of the carbon substrate with and without (Lys)6-tagged rHbI confirmed protein immobilization, as observed by transmission electron microscopy. The electrochemical studies, which were performed to evaluate the redox center of the immobilized protein, show a confinement suitable for an efficient electron transfer system. More importantly, the electrochemical studies allowed determination of a redox potential for the new (Lys)6-tagged rHbI. The data show that the protein is electrochemically active and retains its biological activity toward H2S.},
doi = {10.1021/acsomega.7b01500},
journal = {ACS Omega},
number = 12,
volume = 2,
place = {United States},
year = {Fri Dec 15 00:00:00 EST 2017},
month = {Fri Dec 15 00:00:00 EST 2017}
}
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https://doi.org/10.1021/acsomega.7b01500
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Figures / Tables:

Table 1 Table 1: Immobilized (Lys)6-Tagged rHbI a on MWCNTs-COOH in Control and Coupling Agents' Reactions
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