Alloaffinity filtration: A general approach to the purification of dynein and dynein-like molecules

Anthony Nasr, Peter Satir

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

Alloaffinity filtration simply and specifically separates certain axonemal dyneins and dynein arm components from crude mixtures on the basis of their ability to bind and decorate Tetrahymena axonemal microtubules on a filter in the absence of ATP and to detach and pass into the eluate when 0.5 mm ATP is added. The procedure, which may be performed repetitively, is successful in purifying a Tetrahymena dynein that has characteristics of 30 S dynein prepared by conventional methods, while other dyneins originally present in the mixture, e.g., 14 S Tetrahymena dynein, are not found in the ATP eluate. A relatively homogeneous population of dynein oligomers is obtained. Alloaffinity-purified 30 S Tetrahymena dynein consists of heavy-, intermediate-, and light-chain polypeptides that cosediment in a sucrose gradient in fixed molar ratios and that have structural features of in situ Tetrahymena arms. Dyneins from other species will bind to Tetrahymena microtubules and can be purified by this method. Alloaffinity-purified Chlamydomonas dynein is a set of polypeptides including the four heavy chains that characterize the outer arm.

Original languageEnglish (US)
Pages (from-to)97-108
Number of pages12
JournalAnalytical Biochemistry
Volume151
Issue number1
DOIs
StatePublished - Nov 15 1985

Fingerprint

Dyneins
Purification
Tetrahymena
Molecules
Adenosine Triphosphate
Microtubules
Axonemal Dyneins
Chlamydomonas
Peptides
Oligomers
Sucrose
Light

Keywords

  • ATPase
  • cilia
  • dynein
  • electron microscopy
  • negative stain

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Alloaffinity filtration : A general approach to the purification of dynein and dynein-like molecules. / Nasr, Anthony; Satir, Peter.

In: Analytical Biochemistry, Vol. 151, No. 1, 15.11.1985, p. 97-108.

Research output: Contribution to journalArticle

@article{2d60bde95b224e439ad3370f86197983,
title = "Alloaffinity filtration: A general approach to the purification of dynein and dynein-like molecules",
abstract = "Alloaffinity filtration simply and specifically separates certain axonemal dyneins and dynein arm components from crude mixtures on the basis of their ability to bind and decorate Tetrahymena axonemal microtubules on a filter in the absence of ATP and to detach and pass into the eluate when 0.5 mm ATP is added. The procedure, which may be performed repetitively, is successful in purifying a Tetrahymena dynein that has characteristics of 30 S dynein prepared by conventional methods, while other dyneins originally present in the mixture, e.g., 14 S Tetrahymena dynein, are not found in the ATP eluate. A relatively homogeneous population of dynein oligomers is obtained. Alloaffinity-purified 30 S Tetrahymena dynein consists of heavy-, intermediate-, and light-chain polypeptides that cosediment in a sucrose gradient in fixed molar ratios and that have structural features of in situ Tetrahymena arms. Dyneins from other species will bind to Tetrahymena microtubules and can be purified by this method. Alloaffinity-purified Chlamydomonas dynein is a set of polypeptides including the four heavy chains that characterize the outer arm.",
keywords = "ATPase, cilia, dynein, electron microscopy, negative stain",
author = "Anthony Nasr and Peter Satir",
year = "1985",
month = "11",
day = "15",
doi = "10.1016/0003-2697(85)90058-2",
language = "English (US)",
volume = "151",
pages = "97--108",
journal = "Analytical Biochemistry",
issn = "0003-2697",
publisher = "Academic Press Inc.",
number = "1",

}

TY - JOUR

T1 - Alloaffinity filtration

T2 - A general approach to the purification of dynein and dynein-like molecules

AU - Nasr, Anthony

AU - Satir, Peter

PY - 1985/11/15

Y1 - 1985/11/15

N2 - Alloaffinity filtration simply and specifically separates certain axonemal dyneins and dynein arm components from crude mixtures on the basis of their ability to bind and decorate Tetrahymena axonemal microtubules on a filter in the absence of ATP and to detach and pass into the eluate when 0.5 mm ATP is added. The procedure, which may be performed repetitively, is successful in purifying a Tetrahymena dynein that has characteristics of 30 S dynein prepared by conventional methods, while other dyneins originally present in the mixture, e.g., 14 S Tetrahymena dynein, are not found in the ATP eluate. A relatively homogeneous population of dynein oligomers is obtained. Alloaffinity-purified 30 S Tetrahymena dynein consists of heavy-, intermediate-, and light-chain polypeptides that cosediment in a sucrose gradient in fixed molar ratios and that have structural features of in situ Tetrahymena arms. Dyneins from other species will bind to Tetrahymena microtubules and can be purified by this method. Alloaffinity-purified Chlamydomonas dynein is a set of polypeptides including the four heavy chains that characterize the outer arm.

AB - Alloaffinity filtration simply and specifically separates certain axonemal dyneins and dynein arm components from crude mixtures on the basis of their ability to bind and decorate Tetrahymena axonemal microtubules on a filter in the absence of ATP and to detach and pass into the eluate when 0.5 mm ATP is added. The procedure, which may be performed repetitively, is successful in purifying a Tetrahymena dynein that has characteristics of 30 S dynein prepared by conventional methods, while other dyneins originally present in the mixture, e.g., 14 S Tetrahymena dynein, are not found in the ATP eluate. A relatively homogeneous population of dynein oligomers is obtained. Alloaffinity-purified 30 S Tetrahymena dynein consists of heavy-, intermediate-, and light-chain polypeptides that cosediment in a sucrose gradient in fixed molar ratios and that have structural features of in situ Tetrahymena arms. Dyneins from other species will bind to Tetrahymena microtubules and can be purified by this method. Alloaffinity-purified Chlamydomonas dynein is a set of polypeptides including the four heavy chains that characterize the outer arm.

KW - ATPase

KW - cilia

KW - dynein

KW - electron microscopy

KW - negative stain

UR - http://www.scopus.com/inward/record.url?scp=0022381355&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0022381355&partnerID=8YFLogxK

U2 - 10.1016/0003-2697(85)90058-2

DO - 10.1016/0003-2697(85)90058-2

M3 - Article

C2 - 2936272

AN - SCOPUS:0022381355

VL - 151

SP - 97

EP - 108

JO - Analytical Biochemistry

JF - Analytical Biochemistry

SN - 0003-2697

IS - 1

ER -