Isolation and characterization of multiple molecular forms of cytosolic NAD linked glycerol 3 phosphate dehydrogenase from normal and neoplastic rabbit tissues

M. J. Ostro, Thomas P Fondy

Research output: Contribution to journalArticle

35 Citations (Scopus)

Abstract

Forms of cytosolic NAD-linked glycerol-3-phosphate dehydrogenase were isolated to homogeneity in yields as high as 70% from rabbit muscle, liver, mammary gland, and heart, and partially purified from rabbit Brown Pearce carcinoma. The purified enzyme forms were characterized by flat plate isoelectric focusing, heat inactivation, comparative kinetics, antibody inactivation studies, and amino acid analysis. The enzyme activity in white skeletal muscle, mammary gland, and liver is composed predominantly of a form which focuses at pH 6.5 while the major form from heart focuses at pH 6.1. The major form from muscle, liver, and mammary gland is indistinguishable by all the techniques applied. The heart form of the enzyme is distinct from the major enzyme form in muscle, liver, and mammary gland by flat plate isoelectric focusing, heat inactivation, amino acid analysis, comparative kinetics, and antibody inactivation studies, but has the same subunit molecular weight. Two closely spaced minor forms, isolated from liver, focus between the heart and muscle enzyme forms (pH 6.3). The apparent K(m) values of these forms indicate that they are not hybrids of the heart and muscle isoenzymes. Rabbit Brown Pearce carcinoma glycerol-3-phosphate dehydrogenase was purified by affinity chromatography and appears on flat plate isoelectric focusing as an anionic form distinct from any previously found in normal rabbit tissues.

Original languageEnglish (US)
Pages (from-to)5575-5583
Number of pages9
JournalJournal of Biological Chemistry
Volume252
Issue number15
StatePublished - 1977

Fingerprint

Glycerol-3-Phosphate Dehydrogenase (NAD+)
Glycerolphosphate Dehydrogenase
NAD
Muscle
Human Mammary Glands
Liver
Brown-Pearce Carcinoma
Tissue
Rabbits
Isoelectric Focusing
Enzymes
Muscles
Myocardium
Hot Temperature
Amino Acids
Antibodies
Affinity chromatography
Kinetics
Affinity Chromatography
Isoenzymes

ASJC Scopus subject areas

  • Biochemistry

Cite this

@article{fc42c65eb97143e0aac62bd53f3e680e,
title = "Isolation and characterization of multiple molecular forms of cytosolic NAD linked glycerol 3 phosphate dehydrogenase from normal and neoplastic rabbit tissues",
abstract = "Forms of cytosolic NAD-linked glycerol-3-phosphate dehydrogenase were isolated to homogeneity in yields as high as 70{\%} from rabbit muscle, liver, mammary gland, and heart, and partially purified from rabbit Brown Pearce carcinoma. The purified enzyme forms were characterized by flat plate isoelectric focusing, heat inactivation, comparative kinetics, antibody inactivation studies, and amino acid analysis. The enzyme activity in white skeletal muscle, mammary gland, and liver is composed predominantly of a form which focuses at pH 6.5 while the major form from heart focuses at pH 6.1. The major form from muscle, liver, and mammary gland is indistinguishable by all the techniques applied. The heart form of the enzyme is distinct from the major enzyme form in muscle, liver, and mammary gland by flat plate isoelectric focusing, heat inactivation, amino acid analysis, comparative kinetics, and antibody inactivation studies, but has the same subunit molecular weight. Two closely spaced minor forms, isolated from liver, focus between the heart and muscle enzyme forms (pH 6.3). The apparent K(m) values of these forms indicate that they are not hybrids of the heart and muscle isoenzymes. Rabbit Brown Pearce carcinoma glycerol-3-phosphate dehydrogenase was purified by affinity chromatography and appears on flat plate isoelectric focusing as an anionic form distinct from any previously found in normal rabbit tissues.",
author = "Ostro, {M. J.} and Fondy, {Thomas P}",
year = "1977",
language = "English (US)",
volume = "252",
pages = "5575--5583",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "15",

}

TY - JOUR

T1 - Isolation and characterization of multiple molecular forms of cytosolic NAD linked glycerol 3 phosphate dehydrogenase from normal and neoplastic rabbit tissues

AU - Ostro, M. J.

AU - Fondy, Thomas P

PY - 1977

Y1 - 1977

N2 - Forms of cytosolic NAD-linked glycerol-3-phosphate dehydrogenase were isolated to homogeneity in yields as high as 70% from rabbit muscle, liver, mammary gland, and heart, and partially purified from rabbit Brown Pearce carcinoma. The purified enzyme forms were characterized by flat plate isoelectric focusing, heat inactivation, comparative kinetics, antibody inactivation studies, and amino acid analysis. The enzyme activity in white skeletal muscle, mammary gland, and liver is composed predominantly of a form which focuses at pH 6.5 while the major form from heart focuses at pH 6.1. The major form from muscle, liver, and mammary gland is indistinguishable by all the techniques applied. The heart form of the enzyme is distinct from the major enzyme form in muscle, liver, and mammary gland by flat plate isoelectric focusing, heat inactivation, amino acid analysis, comparative kinetics, and antibody inactivation studies, but has the same subunit molecular weight. Two closely spaced minor forms, isolated from liver, focus between the heart and muscle enzyme forms (pH 6.3). The apparent K(m) values of these forms indicate that they are not hybrids of the heart and muscle isoenzymes. Rabbit Brown Pearce carcinoma glycerol-3-phosphate dehydrogenase was purified by affinity chromatography and appears on flat plate isoelectric focusing as an anionic form distinct from any previously found in normal rabbit tissues.

AB - Forms of cytosolic NAD-linked glycerol-3-phosphate dehydrogenase were isolated to homogeneity in yields as high as 70% from rabbit muscle, liver, mammary gland, and heart, and partially purified from rabbit Brown Pearce carcinoma. The purified enzyme forms were characterized by flat plate isoelectric focusing, heat inactivation, comparative kinetics, antibody inactivation studies, and amino acid analysis. The enzyme activity in white skeletal muscle, mammary gland, and liver is composed predominantly of a form which focuses at pH 6.5 while the major form from heart focuses at pH 6.1. The major form from muscle, liver, and mammary gland is indistinguishable by all the techniques applied. The heart form of the enzyme is distinct from the major enzyme form in muscle, liver, and mammary gland by flat plate isoelectric focusing, heat inactivation, amino acid analysis, comparative kinetics, and antibody inactivation studies, but has the same subunit molecular weight. Two closely spaced minor forms, isolated from liver, focus between the heart and muscle enzyme forms (pH 6.3). The apparent K(m) values of these forms indicate that they are not hybrids of the heart and muscle isoenzymes. Rabbit Brown Pearce carcinoma glycerol-3-phosphate dehydrogenase was purified by affinity chromatography and appears on flat plate isoelectric focusing as an anionic form distinct from any previously found in normal rabbit tissues.

UR - http://www.scopus.com/inward/record.url?scp=0017684404&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0017684404&partnerID=8YFLogxK

M3 - Article

VL - 252

SP - 5575

EP - 5583

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 15

ER -