Late-infantile Batten disease: Purification of the subunit c of the mitochondrial ATP synthase from storage material

Kevork Hagopian, B. D. Lake, B. G. Winchester, J. B. Clark

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

The accumulation of subunit c of the mitochondrial ATP synthase in late- infantile neuronal lipofuscinosis (LINCL) and juvenile neuronal lipofuscinosis (JNCL) is well documented. The purification of the subunit from diverse sources has been reported previously, although not from the brain of Batten disease patients. This proteolipid has now been purified from late-infantile Batten disease brain. The procedures used were an original combination of the conventional solubilisation, differential centrifugation, organic solvent extractions, preparative gel electrophoresis, and FPLC. Gel filtration of the purified protein indicated molecular mass equal to or greater than 2 x 106 Da; however, electrophoresis of this pure protein suggested a molecular mass of approximately 3,500 Da, which is a characteristic of subunit c. The pure protein may be solubilised in aqueous buffer containing <1% lithium dodecyl sulphate (LDS). The protein binds dicyclohexylcarbodiimide (DCCD) and shows immunoreactivity to antibodies raised against ovine storage bodies.

Original languageEnglish (US)
Pages (from-to)272-278
Number of pages7
JournalAmerican Journal of Medical Genetics
Volume57
Issue number2
DOIs
StatePublished - Jun 16 1995
Externally publishedYes

Fingerprint

Mitochondrial Proton-Translocating ATPases
Neuronal Ceroid-Lipofuscinoses
Electrophoresis
Proteins
Dicyclohexylcarbodiimide
Proteolipids
Brain Diseases
Centrifugation
Gel Chromatography
Sheep
Buffers
Gels
Antibodies
Brain

Keywords

  • DCCD
  • late-infantile Batten disease
  • lysosomal storage
  • mitochondrial ATP synthase
  • proteolipid
  • subunit c purification

ASJC Scopus subject areas

  • Genetics(clinical)

Cite this

Late-infantile Batten disease : Purification of the subunit c of the mitochondrial ATP synthase from storage material. / Hagopian, Kevork; Lake, B. D.; Winchester, B. G.; Clark, J. B.

In: American Journal of Medical Genetics, Vol. 57, No. 2, 16.06.1995, p. 272-278.

Research output: Contribution to journalArticle

@article{c7a9cc80865a425aaba0b32f6d04cf9e,
title = "Late-infantile Batten disease: Purification of the subunit c of the mitochondrial ATP synthase from storage material",
abstract = "The accumulation of subunit c of the mitochondrial ATP synthase in late- infantile neuronal lipofuscinosis (LINCL) and juvenile neuronal lipofuscinosis (JNCL) is well documented. The purification of the subunit from diverse sources has been reported previously, although not from the brain of Batten disease patients. This proteolipid has now been purified from late-infantile Batten disease brain. The procedures used were an original combination of the conventional solubilisation, differential centrifugation, organic solvent extractions, preparative gel electrophoresis, and FPLC. Gel filtration of the purified protein indicated molecular mass equal to or greater than 2 x 106 Da; however, electrophoresis of this pure protein suggested a molecular mass of approximately 3,500 Da, which is a characteristic of subunit c. The pure protein may be solubilised in aqueous buffer containing <1{\%} lithium dodecyl sulphate (LDS). The protein binds dicyclohexylcarbodiimide (DCCD) and shows immunoreactivity to antibodies raised against ovine storage bodies.",
keywords = "DCCD, late-infantile Batten disease, lysosomal storage, mitochondrial ATP synthase, proteolipid, subunit c purification",
author = "Kevork Hagopian and Lake, {B. D.} and Winchester, {B. G.} and Clark, {J. B.}",
year = "1995",
month = "6",
day = "16",
doi = "10.1002/ajmg.1320570232",
language = "English (US)",
volume = "57",
pages = "272--278",
journal = "American Journal of Medical Genetics",
issn = "1552-4825",
publisher = "Wiley-Liss Inc.",
number = "2",

}

TY - JOUR

T1 - Late-infantile Batten disease

T2 - Purification of the subunit c of the mitochondrial ATP synthase from storage material

AU - Hagopian, Kevork

AU - Lake, B. D.

AU - Winchester, B. G.

AU - Clark, J. B.

PY - 1995/6/16

Y1 - 1995/6/16

N2 - The accumulation of subunit c of the mitochondrial ATP synthase in late- infantile neuronal lipofuscinosis (LINCL) and juvenile neuronal lipofuscinosis (JNCL) is well documented. The purification of the subunit from diverse sources has been reported previously, although not from the brain of Batten disease patients. This proteolipid has now been purified from late-infantile Batten disease brain. The procedures used were an original combination of the conventional solubilisation, differential centrifugation, organic solvent extractions, preparative gel electrophoresis, and FPLC. Gel filtration of the purified protein indicated molecular mass equal to or greater than 2 x 106 Da; however, electrophoresis of this pure protein suggested a molecular mass of approximately 3,500 Da, which is a characteristic of subunit c. The pure protein may be solubilised in aqueous buffer containing <1% lithium dodecyl sulphate (LDS). The protein binds dicyclohexylcarbodiimide (DCCD) and shows immunoreactivity to antibodies raised against ovine storage bodies.

AB - The accumulation of subunit c of the mitochondrial ATP synthase in late- infantile neuronal lipofuscinosis (LINCL) and juvenile neuronal lipofuscinosis (JNCL) is well documented. The purification of the subunit from diverse sources has been reported previously, although not from the brain of Batten disease patients. This proteolipid has now been purified from late-infantile Batten disease brain. The procedures used were an original combination of the conventional solubilisation, differential centrifugation, organic solvent extractions, preparative gel electrophoresis, and FPLC. Gel filtration of the purified protein indicated molecular mass equal to or greater than 2 x 106 Da; however, electrophoresis of this pure protein suggested a molecular mass of approximately 3,500 Da, which is a characteristic of subunit c. The pure protein may be solubilised in aqueous buffer containing <1% lithium dodecyl sulphate (LDS). The protein binds dicyclohexylcarbodiimide (DCCD) and shows immunoreactivity to antibodies raised against ovine storage bodies.

KW - DCCD

KW - late-infantile Batten disease

KW - lysosomal storage

KW - mitochondrial ATP synthase

KW - proteolipid

KW - subunit c purification

UR - http://www.scopus.com/inward/record.url?scp=0028998942&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0028998942&partnerID=8YFLogxK

U2 - 10.1002/ajmg.1320570232

DO - 10.1002/ajmg.1320570232

M3 - Article

C2 - 7668344

AN - SCOPUS:0028998942

VL - 57

SP - 272

EP - 278

JO - American Journal of Medical Genetics

JF - American Journal of Medical Genetics

SN - 1552-4825

IS - 2

ER -