DsRed as a highly sensitive, selective, and reversible fluorescence-based biosensor for both Cu(+) and Cu(2+) ions.
The wild type form of Red fluorescent protein (DsRed), an intrinsically fluorescent protein found in tropical corals, is found to be highly selective, reversible and sensitive for both Cu(+) and Cu(2+), with a nanomolar detection limit. The selectivity towards these ions is retained even in the presence of other heavy metal ions. The K(d) values for monovalent and divalent copper, based on single binding isotherms, are 450 and 540 nM, respectively. The wild type DsRed sensitivity to Cu(2+) (below 1 ppb) is seven orders of magnitude better than that of the related wild type Green Fluorescent protein (GFP), and it is even 40 times more sensitive than engineered mutants of GFP. Potential binding sites have been proposed, based on amino acid sequences for copper binding and the distance from the chromophore, with the aid of computer modeling.
Sumner
James P
JP
Department of Chemistry, The University of Michigan, 930N University AVE, Ann Arbor, MI 48109, USA.
Westerberg
Nissa M
NM
Stoddard
Andrea K
AK
Hurst
Tamiika K
TK
Cramer
Michele
M
Thompson
Richard B
RB
Fierke
Carol A
CA
Kopelman
Raoul
R
D015374:Biosensing Techniques / Q000295:instrumentation / Q000379:methods*; D003300:Copper / Q000032:analysis* / Q000737:chemistry*; D007477:Ions; D008164:Luminescent Proteins / Q000032:analysis* / Q000737:chemistry*; D013050:Spectrometry, Fluorescence / Q000295:instrumentation / Q000379:methods*
D007477:Ions; D008164:Luminescent Proteins; C413663:fluorescent protein 583; D003300:Copper