Fluorescence lifetime measurements using a total internal reflection fluorimetry

Evidence for a conformational change in albumin adsorbed to quartz

M. R. Rainbow, S. Atherton, R. C. Eberhart

Research output: Contribution to journalArticle

30 Citations (Scopus)

Abstract

We have modified the total internal reflectance fluorimetry (TIRF) method to determine, indirectly, quantum fluorescence yield and thus infer structural changes of adsorbed plasma proteins. We employed a frequency multiplied Nd:YAG pulse laser to excite tryptophan residues of bovine serum albumin (BSA), measuring the resulting fluorescence lifetimes. BSA adsorbed on quartz was investigated at normal pH and under acidic (pH 3. 8) conditions at which albumin shape change has been established in bulk solution. The results support the hypothesis that absorbed albumin exists in the form of a two-layer deposit, each with a possibly different structure. Vigorous washing removes all but the most tightly held protein, which exists in a possibly further unfolded configuration. The decrease in fluorescence lifetime of adsorbed BSA implies a corresponding change in quantum yield. A change in quantum yield of BSA tryptophan fluorophors implies that measuring adsorbed protein concentration by intrinsic fluorescence is unreliable.

Original languageEnglish (US)
Pages (from-to)539-555
Number of pages17
JournalJournal of Biomedical Materials Research
Volume21
Issue number5
StatePublished - May 1987

Fingerprint

Quartz
Bovine Serum Albumin
Albumins
Fluorescence
Quantum yield
Proteins
Tryptophan
Washing
Blood Proteins
Laser pulses
Deposits
Plasmas

ASJC Scopus subject areas

  • Biomedical Engineering
  • Biomaterials

Cite this

Fluorescence lifetime measurements using a total internal reflection fluorimetry : Evidence for a conformational change in albumin adsorbed to quartz. / Rainbow, M. R.; Atherton, S.; Eberhart, R. C.

In: Journal of Biomedical Materials Research, Vol. 21, No. 5, 05.1987, p. 539-555.

Research output: Contribution to journalArticle

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