The identification and sequence of the actin-binding domain of human red blood cell β-spectrin

A. M. Karinch, W. E. Zimmer, Steven Goodman

Research output: Contribution to journalArticle

93 Citations (Scopus)

Abstract

The junctions of the red blood cell membrane skeleton are formed by interactions between spectrin and actin protofilaments. A spectrin tryptic peptide of 16.5-kDa apparent molecular mass (based on sodium dodecyl sulfate-polyacrylamide gel electrophoresis) which binds to F-actin in cosedimentation experiments has been identified. The peptide has been partially purified by gel filtration, anion-, and cation exchange chromatography. Intact spectrin heterodimer causes half-maximal inhibition of the 16.5-kDa peptide/F-actin interaction at a concentration of 5 μM. Comparison of the two-dimensional iodopeptide maps of the 16.5-kDa peptide with maps of α- and β-spectrin, demonstrate that the peptide is generated from the β subunit. It shows no significant relationship to the peptide maps of the β-spectrin domains I-IV. Protein sequencing indicated that this actin-binding domain represents a stretch of amino acids at the N terminus of the β subunit from alanine 47 probably through lysine 186. The sequence derived molecular weight of this actin-binding domain is 16,290 g/mol. The sequence presented represents the region of greatest homology among the spectrin supergene family (spectrin, dystrophin, α-actinin).

Original languageEnglish (US)
Pages (from-to)11833-11840
Number of pages8
JournalJournal of Biological Chemistry
Volume265
Issue number20
StatePublished - 1990
Externally publishedYes

Fingerprint

Spectrin
Actins
Blood
Erythrocytes
Cells
Peptides
Iodoproteins
Actinin
Dystrophin
Protein Sequence Analysis
Molecular mass
Cell membranes
Chromatography
Electrophoresis
Skeleton
Sodium Dodecyl Sulfate
Alanine
Lysine
Gel Chromatography
Anions

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

The identification and sequence of the actin-binding domain of human red blood cell β-spectrin. / Karinch, A. M.; Zimmer, W. E.; Goodman, Steven.

In: Journal of Biological Chemistry, Vol. 265, No. 20, 1990, p. 11833-11840.

Research output: Contribution to journalArticle

@article{509a98ca1aa1471f9df7b8644fbb6a84,
title = "The identification and sequence of the actin-binding domain of human red blood cell β-spectrin",
abstract = "The junctions of the red blood cell membrane skeleton are formed by interactions between spectrin and actin protofilaments. A spectrin tryptic peptide of 16.5-kDa apparent molecular mass (based on sodium dodecyl sulfate-polyacrylamide gel electrophoresis) which binds to F-actin in cosedimentation experiments has been identified. The peptide has been partially purified by gel filtration, anion-, and cation exchange chromatography. Intact spectrin heterodimer causes half-maximal inhibition of the 16.5-kDa peptide/F-actin interaction at a concentration of 5 μM. Comparison of the two-dimensional iodopeptide maps of the 16.5-kDa peptide with maps of α- and β-spectrin, demonstrate that the peptide is generated from the β subunit. It shows no significant relationship to the peptide maps of the β-spectrin domains I-IV. Protein sequencing indicated that this actin-binding domain represents a stretch of amino acids at the N terminus of the β subunit from alanine 47 probably through lysine 186. The sequence derived molecular weight of this actin-binding domain is 16,290 g/mol. The sequence presented represents the region of greatest homology among the spectrin supergene family (spectrin, dystrophin, α-actinin).",
author = "Karinch, {A. M.} and Zimmer, {W. E.} and Steven Goodman",
year = "1990",
language = "English (US)",
volume = "265",
pages = "11833--11840",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "20",

}

TY - JOUR

T1 - The identification and sequence of the actin-binding domain of human red blood cell β-spectrin

AU - Karinch, A. M.

AU - Zimmer, W. E.

AU - Goodman, Steven

PY - 1990

Y1 - 1990

N2 - The junctions of the red blood cell membrane skeleton are formed by interactions between spectrin and actin protofilaments. A spectrin tryptic peptide of 16.5-kDa apparent molecular mass (based on sodium dodecyl sulfate-polyacrylamide gel electrophoresis) which binds to F-actin in cosedimentation experiments has been identified. The peptide has been partially purified by gel filtration, anion-, and cation exchange chromatography. Intact spectrin heterodimer causes half-maximal inhibition of the 16.5-kDa peptide/F-actin interaction at a concentration of 5 μM. Comparison of the two-dimensional iodopeptide maps of the 16.5-kDa peptide with maps of α- and β-spectrin, demonstrate that the peptide is generated from the β subunit. It shows no significant relationship to the peptide maps of the β-spectrin domains I-IV. Protein sequencing indicated that this actin-binding domain represents a stretch of amino acids at the N terminus of the β subunit from alanine 47 probably through lysine 186. The sequence derived molecular weight of this actin-binding domain is 16,290 g/mol. The sequence presented represents the region of greatest homology among the spectrin supergene family (spectrin, dystrophin, α-actinin).

AB - The junctions of the red blood cell membrane skeleton are formed by interactions between spectrin and actin protofilaments. A spectrin tryptic peptide of 16.5-kDa apparent molecular mass (based on sodium dodecyl sulfate-polyacrylamide gel electrophoresis) which binds to F-actin in cosedimentation experiments has been identified. The peptide has been partially purified by gel filtration, anion-, and cation exchange chromatography. Intact spectrin heterodimer causes half-maximal inhibition of the 16.5-kDa peptide/F-actin interaction at a concentration of 5 μM. Comparison of the two-dimensional iodopeptide maps of the 16.5-kDa peptide with maps of α- and β-spectrin, demonstrate that the peptide is generated from the β subunit. It shows no significant relationship to the peptide maps of the β-spectrin domains I-IV. Protein sequencing indicated that this actin-binding domain represents a stretch of amino acids at the N terminus of the β subunit from alanine 47 probably through lysine 186. The sequence derived molecular weight of this actin-binding domain is 16,290 g/mol. The sequence presented represents the region of greatest homology among the spectrin supergene family (spectrin, dystrophin, α-actinin).

UR - http://www.scopus.com/inward/record.url?scp=0025290039&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0025290039&partnerID=8YFLogxK

M3 - Article

VL - 265

SP - 11833

EP - 11840

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 20

ER -