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- Title
Intrinsic tryptophan fluorescence and related energy transfer in Leghemoglobin isolated from Arachis hypogea.
- Authors
Basak, Pijush; Bhattacharyya, Maitree
- Abstract
Objective: To explore the structural properties of heme containing leguminous protein Leghemoglobin (Lb) by means of intrinsic tryptophan (Trp) fluorescence and interaction with fluorescent dye 1-anilinonaphthalene-8-sulphonate (ANS). Hence to compare the melting temperature, quantum yield and energy transfer properties of Lb with other standard globular proteins. Method: Intrinsic Trp fluorescence, extrinsic probe ANS, UV fluorescent signal were used to investigate Förster energy transfer in the proteins. Quantum yield and melting temperature (Tm) were determined to characterize Lb isolated from Arachis hypogea. Result: Binding of ANS with the proteins Bovine serum albumin (BSA), lysozyme, Ovalbumin (Oval) and Cytochrome-C (Cyt-C) manifested differential enhancement of ANS fluorescence revealing energy transfer from Trp. Förster equation was used to estimate the efficiency of energy transfer from Trp to ANS. Estimated binding constants were different for 295 nm and 375 nm excitation suggesting the involvement of an additional pathway in Lb via Förster resonance energy transfer (FRET). This is reflected in higher KD value indicating lower binding affinity of ANS-Lb. Conclusion: This is a pioneering endeavor to unfold the structural properties of Lb isolated from Arachis hypogea, since there is no report regarding the spectroscopic properties of this protein, which is of immense agricultural importance. Our work revealed a comparison of thermal stability of heme containing globular proteins which followed the order: Hb > Lb > Cyt-C. Quantum yield and the binding constant for Trp-ANS interaction of Lb were determined. Apparent distance for Trp-ANS energy transfer in Lb and other globular proteins were explored to follow the order: Oval < Lb < BSA < Cyt-C < Lysozyme.
- Subjects
LEGHEMOGLOBIN; TRYPTOPHAN; ANILINONAPHTHALENE sulfonate; TEMPERATURE; ENERGY transfer; PEANUT research; SERUM albumin; CYTOCHROME c
- Publication
Turkish Journal of Biochemistry / Turk Biyokimya Dergisi, 2013, Vol 38, Issue 1, p49
- ISSN
0250-4685
- Publication type
Article
- DOI
10.5505/tjb.2013.53315