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dc.contributor.authorKamal, J K Amisha-
dc.contributor.authorBehere, Digambar V-
dc.description.abstractThe binding of monomeric heme to human serum albumin (HSA) was investigated using steady-state fluorescence, circular dichroism (CD) and optical difference spectroscopic (ODS) techniques. The existence of one strong binding site for heme on HSA was confirmed by titrating heme with HSA and following the quenching of tryptophan (Trp214) fluorescence emission intensity that occurred due to energy transfer. Up to around 1:1 stoichiometric ratio of HSA/heme, the quenching was observed to be very strong, however at higher ratios the quenching progressed very weakly. Similarly, the negative CD band centered at ~397 nm, which appeared on adding heme to HSA, increased in intensity on sequential addition of heme up to [heme]/[HSA]=1. Titration of HSA with heme was followed by ODS and the dissociation constant KD = (4.0±1.0)×10⁻⁵ M was deduced. Results have been explained on the basis of Michaelis-Menton type of mechanism for the heme binding, in which heme first binds reversibly to His146 at the surface of the protein to form an intermediate complex, followed by irreversible binding to Tyr161 in the interior of the proteinen_US
dc.relation.ispartofseriesG01J 3/00en_US
dc.sourceIJBB Vol.42(1) [February 2005]en_US
dc.subjecthuman serum albumin (HSA)en_US
dc.subjectmethemalbumin (MHA)en_US
dc.subjectsteady-state fluorescenceen_US
dc.subjectcircular dichroismen_US
dc.subjectoptical difference spectroscopyen_US
dc.subjecttwo stage bindingen_US
dc.titleBinding of heme to human serum albumin: Steady-state fluorescence, circular dichroism and optical difference spectroscopic studiesen_US
Appears in Collections:IJBB Vol.42(1) [February 2005]

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