PMID- 10407156
OWN - NLM
STAT- MEDLINE
DCOM- 19990820
LR  - 20190610
IS  - 0006-3002 (Print)
IS  - 0006-3002 (Linking)
VI  - 1432
IP  - 2
DP  - 1999 Jul 13
TI  - Nucleotide-dependent bisANS binding to tubulin.
PG  - 350-5
AB  - Non-covalent hydrophobic probes such as 5, 
      5'-bis(8-anilino-1-naphthalenesulfonate) (bisANS) have become increasingly 
      popular to gain information about protein structure and conformation. However, 
      there are limitations as bisANS binds non-specifically at multiple sites of many 
      proteins. Successful use of this probe depends upon the development of binding 
      conditions where only specific dye-protein interaction will occur. In this 
      report, we have shown that the binding of bisANS to tubulin occurs 
      instantaneously, specifically at one high affinity site when 1 mM guanosine 
      5'-triphosphate (GTP) is included in the reaction medium. Substantial portions of 
      protein secondary structure and colchicine binding activity of tubulin are lost 
      upon bisANS binding in absence of GTP. BisANS binding increases with time and 
      occurs at multiple sites in the absence of GTP. Like GTP, other analogs, 
      guanosine 5'-diphosphate, guanosine 5'-monophosphate and adenosine 
      5'-triphosphate, also displace bisANS from the lower affinity sites of tubulin. 
      We believe that these multiple binding sites are generated due to the 
      bisANS-induced structural changes on tubulin and the presence of GTP and other 
      nucleotides protect those structural changes.
FAU - Chakraborty, S
AU  - Chakraborty S
AD  - Department of Biochemistry, Bose Institute, Centenary Building, P-1/12 C.I.T. 
      Scheme VII M, Calcutta 700054, India.
FAU - Sarkar, N
AU  - Sarkar N
FAU - Bhattacharyya, B
AU  - Bhattacharyya B
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - Netherlands
TA  - Biochim Biophys Acta
JT  - Biochimica et biophysica acta
JID - 0217513
RN  - 0 (Anilino Naphthalenesulfonates)
RN  - 0 (Tubulin)
RN  - 63741-13-9 (5,5'-bis(8-(phenylamino)-1-naphthalenesulfonate))
RN  - 86-01-1 (Guanosine Triphosphate)
RN  - SML2Y3J35T (Colchicine)
SB  - IM
MH  - Anilino Naphthalenesulfonates/*chemistry
MH  - Circular Dichroism
MH  - Colchicine/chemistry
MH  - Fluorescence
MH  - Guanosine Triphosphate/chemistry
MH  - Kinetics
MH  - Temperature
MH  - Tubulin/*chemistry
EDAT- 1999/07/17 00:00
MHDA- 1999/07/17 00:01
CRDT- 1999/07/17 00:00
PHST- 1999/07/17 00:00 [pubmed]
PHST- 1999/07/17 00:01 [medline]
PHST- 1999/07/17 00:00 [entrez]
AID - S0167-4838(99)00110-7 [pii]
AID - 10.1016/s0167-4838(99)00110-7 [doi]
PST - ppublish
SO  - Biochim Biophys Acta. 1999 Jul 13;1432(2):350-5. doi: 
      10.1016/s0167-4838(99)00110-7.