fulltext.study @t Gmail

Photocleavage of avidin by a new pyrenyl probe

Paper ID Volume ID Publish Year Pages File Format Full-Text
29678 44429 2011 5 PDF Available
Title
Photocleavage of avidin by a new pyrenyl probe
Abstract

In this study, a new small-molecule-based reagent was designed to recognize and bind to specific site in protein. A new pyrenyl probe, d-biotinyl-1(1-pyrene)methylamide (Py–biotin) was designed and synthesized by coupling of d-biotin to 1(1-pyrene)methylamine hydrochloride. Binding studies and site-specific photocleavage of avidin by Py–biotin were demonstrated. Binding of Py–biotin to avidin was studied using absorbance and fluorescence spectroscopic techniques. Red shifts of the absorption peak positions of the pyrenyl chromophore followed by hyperchromism were observed upon binding to avidin. The photocleavage of avidin was achieved when a mixture of the protein, Py–biotin, and an electron acceptor, cobalt(III) hexammine trichloride (CoHA), was irradiated at 342 nm. No reaction occurred in the absence of the probe, CoHA, or light. N-terminal sequencing of the peptide fragments indicated a cleavage site of avidin between Thr 77 and Val 78. The high specificity of photocleavage may be valuable in targeting specific sites of proteins with small molecules.

Graphical abstractA new small reagent, d-biotinyl-1(1-pyrene)methylamide (Py–biotin), was designed to bind to specific site of avidin. Site-specific photocleavage of avidin was successfully achieved upon irradiation of avidin/Py–biotin solution in the presence of an electron acceptor, CoHA.Figure optionsDownload full-size imageDownload as PowerPoint slideHighlights► Avidin was successfully cleaved at specific binding site by the new probe, Py–biotin. ► Two new cleaved fragments were obtained with cleavage site between Thr 77–Val 78. ► Photocleavage cannot be achieved in the absence of either electron acceptor, Py–biotin or light.

Keywords
Pyrene; Photocleavage; Fluorescence quenching; Avidin; Biotin
First Page Preview
Photocleavage of avidin by a new pyrenyl probe
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us
Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Photochemistry and Photobiology B: Biology - Volume 103, Issue 3, 2 June 2011, Pages 251–255
Authors
, , , ,
Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us