NDLI: Cyclic AMP Receptor Protein-Aequorin Molecular Switch for Cyclic AMP

Content Provider	PubMed Central
Author	Scott, Daniel Teasley, Hamorsky Krystal Ensor, C. Mark Anderson, Kimberly W. Sylvia, Daunert
Abstract	Molecular switches are designer molecules that combine the functionality of two individual proteins into one, capable of manifesting an “on/off” signal in response to a stimulus. These switches have unique properties and functionalities and thus, can be employed as nanosensors in a variety of applications. To that end, we have developed a bioluminescent molecular switch for cyclic AMP. Bioluminescence offers many advantages over fluorescence and other detection methods including the fact that there is essentially zero background signal in physiological fluids, allowing for more sensitive detection and monitoring. The switch was created by combining the properties of the cyclic AMP receptor protein (CRP), a transcriptional regulatory protein from E. coli that binds selectively to cAMP with those of aequorin, a bioluminescent photoprotein native of the jellyfish Aequorea victoria. Genetic manipulation to split the genetic coding sequence of aequorin in two and genetically attach the fragments to the N and C termini of CRP, resulted in a hybrid protein molecular switch. The conformational change experienced by CRP upon the binding of cyclic AMP is suspected to result in the observed loss of bioluminescent signal from aequorin. The “on/off” bioluminescence can be modulated by cyclic AMP over a range of several orders of magnitude in a linear fashion in addition to the capacity to detect changes in cellular cyclic AMP of intact cells exposed to different external stimuli without the need to lyse the cells. We envision that the molecular switch could find applications in vitro as well as in vivo cyclic AMP detection and/or imaging.
Related Links	http://dx.doi.org/10.1021/bc100486b
Ending Page	481
Page Count	7
Starting Page	475
File Format	PDF
ISSN	10431802
e-ISSN	15204812
Journal	Bioconjugate chemistry
Issue Number	3
Volume Number	22
Language	English
Publisher Date	2011-03-16
Access Restriction	Open
Subject Keyword	Biotechnology Organic Chemistry Bioengineering Pharmacology Pharmaceutical Science Biomedical Engineering Research in Higher Education
Content Type	Text
Resource Type	Article
Subject	Organic Chemistry Pharmacology Bioengineering Biomedical Engineering Biotechnology Pharmaceutical Science

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1	Ministry of Education (GoI), Department of Higher Education	Sanctioning Authority	https://www.education.gov.in/ict-initiatives
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3	National Digital Library of India Office, Indian Institute of Technology Kharagpur	The administrative and infrastructural headquarters of the project	Dr. B. Sutradhar bsutra@ndl.gov.in
4	Project PI / Joint PI	Principal Investigator and Joint Principal Investigators of the project	Dr. B. Sutradhar bsutra@ndl.gov.in Prof. Saswat Chakrabarti will be added soon
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