Fluorescence studies of human semi-beta-hemoglobin assembly.

Abstract:

:The intrinsic fluorescence properties of human alpha apohemoglobin at protein concentrations from 1 to 5 microM in 0.1 M potassium phosphate buffer, pH 7 or 8 at 5 degrees C were monitored in the absence and presence of a fixed concentration (5 microM) of a fluorescence quenching heme-containing native or Des (146-His, 145-Tyr) beta chain partner. These "reverse quenching" studies revealed that the emission intensity changes observed correlated well with protein concentration and theoretical extent of semi-beta-hemoglobin assembly. Furthermore, the relative quenching efficiencies were calculated to be 0.32, 0.25 and 0.61 for beta (pH 7), beta (pH 8) and Des beta (pH 7) chains, respectively. Thus, heme-mediated quenching was sensitive to the expected pH induced alpha apohemoglobin conformational change and to alteration in beta chain structure. Intramolecular changes induced by carboxylterminal modification (decreased "beta chain self-quenching") appeared to enhance the intermolecular rearrangements (increased "alpha chain partner quenching") seen upon subunit assembly.

authors

Chiu F,Vasudevan G,Morris A,McDonald MJ

doi

10.1006/bbrc.1997.7955

subject

Has Abstract

pub_date

1998-01-14 00:00:00

pages

365-8

issue

2

eissn

0006-291X

issn

1090-2104

pii

S0006-291X(97)97955-9

journal_volume

242

pub_type

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