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Probing the nucleotide binding sites in T7 RNA polymerase using cibacron blue

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Title Probing the nucleotide binding sites in T7 RNA polymerase using cibacron blue
 
Creator Das, Mili
Dasgupta, Dipak
 
Description 78-83
T7 RNA polymerase (T7 RNAP) is an
enzyme that utilizes ribonucleotides to synthesize the nascent RNA chain in a
template-dependent manner. In this work we have studied the interaction of T7
RNAP with cibacron blue, an anthraquinone monochlorotriazine dye, and its
effect on the function of the enzyme. T7 RNAP binds to the dye in a bi-phasic
manner. The first phase of the binding is characterized by a high affinity (Kd
in the nanomolar range) and reversible inactivation of the enzyme. The
second binding site is the common substrate binding site. The association of
the dye with T7 RNAP is a good model to understand the physiological
significance of a high affinity binding of the initiating nucleotide, GTP,
earlier reported from our laboratory. The results will be discussed to
understand the role of the high affinity GTP binding.
 
Date 2013-07-16T06:10:18Z
2013-07-16T06:10:18Z
2001-04
 
Type Article
 
Identifier 0975-0959 (Online); 0301-1208 (Print)
http://hdl.handle.net/123456789/19803
 
Language en_US
 
Rights CC Attribution-Noncommercial-No Derivative Works 2.5 India
 
Publisher NISCAIR-CSIR, India
 
Source IJBB Vol.38(1-2) [February-April 2001]