The mode of chaperoning of dithiothreitol-denatured α-lactalbumin by α-crystallin

Frederick A. Bettelheim, Rafat Ansari, Qiu Fang Cheng, J. Samuel Zigler

Research output: Contribution to journalArticle

Abstract

Molecular chaperones prevent the aggregation of partially folded or misfolded forms of protein. α-crystallin performs such a function in the ocular lens. To gain insight into the mechanism of the anti-aggregation activity of α-crystallin, we performed dynamic light scattering (DLS) measurements investigating its interaction with partially denatured α-lactalbumin over a 24 hr period. Analyses were conducted as a function of the concentration of α-lactalbumin as well as the bovine α-crystallin/α-lactalbumin ratio. Additional studies of the systems were performed by HPLC and SDS gel electrophoresis. The particle distribution patterns derived from the DLS data indicated that the chaperoned complex (lactalbumin plus crystallin) is a loose fluffy globular entity. After the complex becomes saturated with lactalbumin, it appears to release the partially denatured lactalbumin which may aggregate into high molecular weight moieties. These eventually may precipitate out of solution. On longer standing, 24 hr and over, the chaperoned complex as well as the lactalbumin aggregates become more compact. The chaperoned complex (α-crystallin plus α-lactalbumin) is in dynamic equilibrium both with the monomeric and the aggregated α-lactalbumin population.

Original languageEnglish (US)
Pages (from-to)292-297
Number of pages6
JournalBiochemical and Biophysical Research Communications
Volume261
Issue number2
DOIs
StatePublished - Aug 2 1999
Externally publishedYes

Fingerprint

Lactalbumin
Crystallins
Dithiothreitol
Dynamic light scattering
Agglomeration
Crystalline Lens
Molecular Chaperones
Electrophoresis
Precipitates
Lenses
Molecular Weight
Gels
Molecular weight
High Pressure Liquid Chromatography

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

The mode of chaperoning of dithiothreitol-denatured α-lactalbumin by α-crystallin. / Bettelheim, Frederick A.; Ansari, Rafat; Cheng, Qiu Fang; Zigler, J. Samuel.

In: Biochemical and Biophysical Research Communications, Vol. 261, No. 2, 02.08.1999, p. 292-297.

Research output: Contribution to journalArticle

Bettelheim, Frederick A. ; Ansari, Rafat ; Cheng, Qiu Fang ; Zigler, J. Samuel. / The mode of chaperoning of dithiothreitol-denatured α-lactalbumin by α-crystallin. In: Biochemical and Biophysical Research Communications. 1999 ; Vol. 261, No. 2. pp. 292-297.
@article{fa030525fa5146d4a0a967066c01aeb0,
title = "The mode of chaperoning of dithiothreitol-denatured α-lactalbumin by α-crystallin",
abstract = "Molecular chaperones prevent the aggregation of partially folded or misfolded forms of protein. α-crystallin performs such a function in the ocular lens. To gain insight into the mechanism of the anti-aggregation activity of α-crystallin, we performed dynamic light scattering (DLS) measurements investigating its interaction with partially denatured α-lactalbumin over a 24 hr period. Analyses were conducted as a function of the concentration of α-lactalbumin as well as the bovine α-crystallin/α-lactalbumin ratio. Additional studies of the systems were performed by HPLC and SDS gel electrophoresis. The particle distribution patterns derived from the DLS data indicated that the chaperoned complex (lactalbumin plus crystallin) is a loose fluffy globular entity. After the complex becomes saturated with lactalbumin, it appears to release the partially denatured lactalbumin which may aggregate into high molecular weight moieties. These eventually may precipitate out of solution. On longer standing, 24 hr and over, the chaperoned complex as well as the lactalbumin aggregates become more compact. The chaperoned complex (α-crystallin plus α-lactalbumin) is in dynamic equilibrium both with the monomeric and the aggregated α-lactalbumin population.",
author = "Bettelheim, {Frederick A.} and Rafat Ansari and Cheng, {Qiu Fang} and Zigler, {J. Samuel}",
year = "1999",
month = "8",
day = "2",
doi = "10.1006/bbrc.1999.1031",
language = "English (US)",
volume = "261",
pages = "292--297",
journal = "Biochemical and Biophysical Research Communications",
issn = "0006-291X",
publisher = "Academic Press Inc.",
number = "2",

}

TY - JOUR

T1 - The mode of chaperoning of dithiothreitol-denatured α-lactalbumin by α-crystallin

AU - Bettelheim, Frederick A.

AU - Ansari, Rafat

AU - Cheng, Qiu Fang

AU - Zigler, J. Samuel

PY - 1999/8/2

Y1 - 1999/8/2

N2 - Molecular chaperones prevent the aggregation of partially folded or misfolded forms of protein. α-crystallin performs such a function in the ocular lens. To gain insight into the mechanism of the anti-aggregation activity of α-crystallin, we performed dynamic light scattering (DLS) measurements investigating its interaction with partially denatured α-lactalbumin over a 24 hr period. Analyses were conducted as a function of the concentration of α-lactalbumin as well as the bovine α-crystallin/α-lactalbumin ratio. Additional studies of the systems were performed by HPLC and SDS gel electrophoresis. The particle distribution patterns derived from the DLS data indicated that the chaperoned complex (lactalbumin plus crystallin) is a loose fluffy globular entity. After the complex becomes saturated with lactalbumin, it appears to release the partially denatured lactalbumin which may aggregate into high molecular weight moieties. These eventually may precipitate out of solution. On longer standing, 24 hr and over, the chaperoned complex as well as the lactalbumin aggregates become more compact. The chaperoned complex (α-crystallin plus α-lactalbumin) is in dynamic equilibrium both with the monomeric and the aggregated α-lactalbumin population.

AB - Molecular chaperones prevent the aggregation of partially folded or misfolded forms of protein. α-crystallin performs such a function in the ocular lens. To gain insight into the mechanism of the anti-aggregation activity of α-crystallin, we performed dynamic light scattering (DLS) measurements investigating its interaction with partially denatured α-lactalbumin over a 24 hr period. Analyses were conducted as a function of the concentration of α-lactalbumin as well as the bovine α-crystallin/α-lactalbumin ratio. Additional studies of the systems were performed by HPLC and SDS gel electrophoresis. The particle distribution patterns derived from the DLS data indicated that the chaperoned complex (lactalbumin plus crystallin) is a loose fluffy globular entity. After the complex becomes saturated with lactalbumin, it appears to release the partially denatured lactalbumin which may aggregate into high molecular weight moieties. These eventually may precipitate out of solution. On longer standing, 24 hr and over, the chaperoned complex as well as the lactalbumin aggregates become more compact. The chaperoned complex (α-crystallin plus α-lactalbumin) is in dynamic equilibrium both with the monomeric and the aggregated α-lactalbumin population.

UR - http://www.scopus.com/inward/record.url?scp=0033517042&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0033517042&partnerID=8YFLogxK

U2 - 10.1006/bbrc.1999.1031

DO - 10.1006/bbrc.1999.1031

M3 - Article

C2 - 10425180

AN - SCOPUS:0033517042

VL - 261

SP - 292

EP - 297

JO - Biochemical and Biophysical Research Communications

JF - Biochemical and Biophysical Research Communications

SN - 0006-291X

IS - 2

ER -