Optical sensing systems based on biomolecular recognition of recombinant proteins

Lyndon L.E. Salins; Vesna Schauer-Vukasinovic; Sylvia Daunert

doi:10.1117/12.308378

15 May 1998 Optical sensing systems based on biomolecular recognition of recombinant proteins

Lyndon L.E. Salins, Vesna Schauer-Vukasinovic, Sylvia Daunert

Proceedings Volume 3270, Methods for Ultrasensitive Detection; (1998) https://doi.org/10.1117/12.308378
Event: Optoelectronics and High-Power Lasers and Applications, 1998, San Jose, CA, United States

Abstract

SIte-directed mutagenesis and the associated site-specific fluorescent labeling of proteins can be used to rationally design reagentless fluorescent molecular senors. The phosphate binding protein (PBP) and calmodulin (CaM) bind to phosphate and calcium in a highly specific manner. These ions induce a hinge motion in the proteins, and the resultant conformational change constitutes the basis of the sensor development. By labeling each protein at a specific site with environment-sensitive fluorescent probes, these conformational changes can be monitored and related to the amount of analyte ion present. In this study, the polymerase chain reaction was used to construct mutants of PBP and CaM that have a single cysteine at positions 197 and 109, respectively. Each protein was site-specifically labeled through the sulfhydryl group of the introduced cysteine residue at a single location with an environment-sensitive fluorescent probe. Characterization of the steady-state fluorescence indicated an enhancement of signal intensity upon binding of the analyte ion. Highly sensitive and selective and selective sensing systems for phosphate and calcium were obtained by using this approach.

Citation Download Citation

Lyndon L.E. Salins, Vesna Schauer-Vukasinovic, and Sylvia Daunert "Optical sensing systems based on biomolecular recognition of recombinant proteins", Proc. SPIE 3270, Methods for Ultrasensitive Detection, (15 May 1998); https://doi.org/10.1117/12.308378

ACCESS THE FULL ARTICLE

INSTITUTIONAL
Select your institution to access the SPIE Digital Library.

SELECT YOUR INSTITUTION

PERSONAL
Sign in with your SPIE account to access your personal subscriptions or to use specific features such as save to my library, sign up for alerts, save searches, etc.

PERSONAL SIGN IN

No SPIE Account? Create one

PURCHASE THIS CONTENT

SUBSCRIBE TO DIGITAL LIBRARY

50 downloads per 1-year subscription

Members: $195

Non-members: $335 ADD TO CART

25 downloads per 1 - year subscription

Members: $145

Non-members: $250 ADD TO CART

PURCHASE SINGLE ARTICLE

Includes PDF, HTML & Video, when available