Heterogeneity in the human erythrocyte Band 3 anion-transporter revealed by Triton X-114 phase partitioning

M. L. Swanson, R. K. Keast, M. L. Jennings, Jeffrey E. Pessin

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

Triton X-114 phase partitioning used in conjunction with countercurrent distribution was utilized to examine the phasing properties of the human erythrocyte Band 3 anion-transport protein. Phase partitioning and countercurrent distribution of Band 3 protein followed by electrophoresis and immunoblotting revealed that Band 3 protein possesses biphasic properties with approx. 65% of the Band 3 97,000-M(r) species being localized in the detergent phase and 35% isolated in the aqueous phase. The bidirectional phasing of the anion-transporter does not appear to be a result of glycosylation or phosphorylation, since treatment of alkali-washed ghosts with glycosidases or phosphatase respectively did not significantly alter the phasing profiles. Chymotrypsin treatment of erythrocytes followed by the purification of the 60,000-M(r) fragment, and exposure of this fragment to phase separation and countercurrent distribution also revealed biphasic partitioning with 70% of the species being isolated in the aqueous phase and 30% in the detergent phase. These data demonstrate that the human erythrocyte Band 3 anion-transport protein is heterogeneous by Triton X-114 phase partitioning and that this heterogeneity is preserved in the 60,000-M(r) chymotryptic fragment of Band 3 protein.

Original languageEnglish (US)
Pages (from-to)229-234
Number of pages6
JournalBiochemical Journal
Volume255
Issue number1
StatePublished - 1988
Externally publishedYes

Fingerprint

Countercurrent Distribution
Erythrocyte Anion Exchange Protein 1
Anions
Erythrocytes
Detergents
Glycoside Hydrolases
Chymotrypsin
Alkalies
Glycosylation
Phosphoric Monoester Hydrolases
Immunoblotting
Electrophoresis
Phosphorylation
Phase separation
Purification
Nonidet P-40
human SLC4A1 protein

ASJC Scopus subject areas

  • Biochemistry

Cite this

Heterogeneity in the human erythrocyte Band 3 anion-transporter revealed by Triton X-114 phase partitioning. / Swanson, M. L.; Keast, R. K.; Jennings, M. L.; Pessin, Jeffrey E.

In: Biochemical Journal, Vol. 255, No. 1, 1988, p. 229-234.

Research output: Contribution to journalArticle

@article{86d7e4172ae247dfaa811c5fdf516697,
title = "Heterogeneity in the human erythrocyte Band 3 anion-transporter revealed by Triton X-114 phase partitioning",
abstract = "Triton X-114 phase partitioning used in conjunction with countercurrent distribution was utilized to examine the phasing properties of the human erythrocyte Band 3 anion-transport protein. Phase partitioning and countercurrent distribution of Band 3 protein followed by electrophoresis and immunoblotting revealed that Band 3 protein possesses biphasic properties with approx. 65{\%} of the Band 3 97,000-M(r) species being localized in the detergent phase and 35{\%} isolated in the aqueous phase. The bidirectional phasing of the anion-transporter does not appear to be a result of glycosylation or phosphorylation, since treatment of alkali-washed ghosts with glycosidases or phosphatase respectively did not significantly alter the phasing profiles. Chymotrypsin treatment of erythrocytes followed by the purification of the 60,000-M(r) fragment, and exposure of this fragment to phase separation and countercurrent distribution also revealed biphasic partitioning with 70{\%} of the species being isolated in the aqueous phase and 30{\%} in the detergent phase. These data demonstrate that the human erythrocyte Band 3 anion-transport protein is heterogeneous by Triton X-114 phase partitioning and that this heterogeneity is preserved in the 60,000-M(r) chymotryptic fragment of Band 3 protein.",
author = "Swanson, {M. L.} and Keast, {R. K.} and Jennings, {M. L.} and Pessin, {Jeffrey E.}",
year = "1988",
language = "English (US)",
volume = "255",
pages = "229--234",
journal = "Biochemical Journal",
issn = "0264-6021",
publisher = "Portland Press Ltd.",
number = "1",

}

TY - JOUR

T1 - Heterogeneity in the human erythrocyte Band 3 anion-transporter revealed by Triton X-114 phase partitioning

AU - Swanson, M. L.

AU - Keast, R. K.

AU - Jennings, M. L.

AU - Pessin, Jeffrey E.

PY - 1988

Y1 - 1988

N2 - Triton X-114 phase partitioning used in conjunction with countercurrent distribution was utilized to examine the phasing properties of the human erythrocyte Band 3 anion-transport protein. Phase partitioning and countercurrent distribution of Band 3 protein followed by electrophoresis and immunoblotting revealed that Band 3 protein possesses biphasic properties with approx. 65% of the Band 3 97,000-M(r) species being localized in the detergent phase and 35% isolated in the aqueous phase. The bidirectional phasing of the anion-transporter does not appear to be a result of glycosylation or phosphorylation, since treatment of alkali-washed ghosts with glycosidases or phosphatase respectively did not significantly alter the phasing profiles. Chymotrypsin treatment of erythrocytes followed by the purification of the 60,000-M(r) fragment, and exposure of this fragment to phase separation and countercurrent distribution also revealed biphasic partitioning with 70% of the species being isolated in the aqueous phase and 30% in the detergent phase. These data demonstrate that the human erythrocyte Band 3 anion-transport protein is heterogeneous by Triton X-114 phase partitioning and that this heterogeneity is preserved in the 60,000-M(r) chymotryptic fragment of Band 3 protein.

AB - Triton X-114 phase partitioning used in conjunction with countercurrent distribution was utilized to examine the phasing properties of the human erythrocyte Band 3 anion-transport protein. Phase partitioning and countercurrent distribution of Band 3 protein followed by electrophoresis and immunoblotting revealed that Band 3 protein possesses biphasic properties with approx. 65% of the Band 3 97,000-M(r) species being localized in the detergent phase and 35% isolated in the aqueous phase. The bidirectional phasing of the anion-transporter does not appear to be a result of glycosylation or phosphorylation, since treatment of alkali-washed ghosts with glycosidases or phosphatase respectively did not significantly alter the phasing profiles. Chymotrypsin treatment of erythrocytes followed by the purification of the 60,000-M(r) fragment, and exposure of this fragment to phase separation and countercurrent distribution also revealed biphasic partitioning with 70% of the species being isolated in the aqueous phase and 30% in the detergent phase. These data demonstrate that the human erythrocyte Band 3 anion-transport protein is heterogeneous by Triton X-114 phase partitioning and that this heterogeneity is preserved in the 60,000-M(r) chymotryptic fragment of Band 3 protein.

UR - http://www.scopus.com/inward/record.url?scp=0023747594&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0023747594&partnerID=8YFLogxK

M3 - Article

C2 - 3196316

AN - SCOPUS:0023747594

VL - 255

SP - 229

EP - 234

JO - Biochemical Journal

JF - Biochemical Journal

SN - 0264-6021

IS - 1

ER -