Uptake and degradation of glyceraldehyde-3-phosphate dehydrogenase by rat liver lysosomes

Fernando Aniento, Enrique Roche, Ana Maria Cuervo, Erwin Knecht

Research output: Contribution to journalArticle

116 Citations (Scopus)

Abstract

The molecular mechanisms involved in the degradation of individual cellular proteins are probably unique and characteristic. We have investigated in rat liver the degradation of glyceraldehyde-3-phosphate dehydrogenase, an abundant cytosolic enzyme of the glycolytic pathway. Immunoblot analysis of isolated liver lysosomes from rats treated with lysosomal inhibitors show that this protein is degraded, at least in part, by a lysosomal pathway. This pathway was further investigated by incubating the enzyme with lysosomes in a cell-free system, followed by proteolysis measurements, sodium dodecyl sulfate-polyacrylamide gel electrophoresis of lysosomes, and electron microscopic immunocytochemistry. We postulate that the degradative mechanism of glyceraldehyde-3-phosphate dehydrogenase includes a temperature-dependent lysosomal pathway, different from classical nonspecific macroautophagy. The postulated pathway involves: binding of the enzyme to the lysosomal membrane, entry into the lysosomal matrix, and degradation. This cell-free system, which can also incorporate in vitro synthesized proteins, should allow further advances toward clarifying the complex signals that regulate protein degradation as well as its close interrelationship with protein synthesis.

Original languageEnglish (US)
Pages (from-to)10463-10470
Number of pages8
JournalJournal of Biological Chemistry
Volume268
Issue number14
StatePublished - May 15 1993
Externally publishedYes

Fingerprint

Glyceraldehyde-3-Phosphate Dehydrogenases
Lysosomes
Liver
Rats
Degradation
Cell-Free System
Proteolysis
Proteins
Enzymes
Autophagy
Sodium Dodecyl Sulfate
Polyacrylamide Gel Electrophoresis
Electrophoresis
Immunohistochemistry
Electrons
Temperature
Membranes

ASJC Scopus subject areas

  • Biochemistry

Cite this

Uptake and degradation of glyceraldehyde-3-phosphate dehydrogenase by rat liver lysosomes. / Aniento, Fernando; Roche, Enrique; Cuervo, Ana Maria; Knecht, Erwin.

In: Journal of Biological Chemistry, Vol. 268, No. 14, 15.05.1993, p. 10463-10470.

Research output: Contribution to journalArticle

Aniento, Fernando ; Roche, Enrique ; Cuervo, Ana Maria ; Knecht, Erwin. / Uptake and degradation of glyceraldehyde-3-phosphate dehydrogenase by rat liver lysosomes. In: Journal of Biological Chemistry. 1993 ; Vol. 268, No. 14. pp. 10463-10470.
@article{7513ee3028d347568f2264b39fe2f378,
title = "Uptake and degradation of glyceraldehyde-3-phosphate dehydrogenase by rat liver lysosomes",
abstract = "The molecular mechanisms involved in the degradation of individual cellular proteins are probably unique and characteristic. We have investigated in rat liver the degradation of glyceraldehyde-3-phosphate dehydrogenase, an abundant cytosolic enzyme of the glycolytic pathway. Immunoblot analysis of isolated liver lysosomes from rats treated with lysosomal inhibitors show that this protein is degraded, at least in part, by a lysosomal pathway. This pathway was further investigated by incubating the enzyme with lysosomes in a cell-free system, followed by proteolysis measurements, sodium dodecyl sulfate-polyacrylamide gel electrophoresis of lysosomes, and electron microscopic immunocytochemistry. We postulate that the degradative mechanism of glyceraldehyde-3-phosphate dehydrogenase includes a temperature-dependent lysosomal pathway, different from classical nonspecific macroautophagy. The postulated pathway involves: binding of the enzyme to the lysosomal membrane, entry into the lysosomal matrix, and degradation. This cell-free system, which can also incorporate in vitro synthesized proteins, should allow further advances toward clarifying the complex signals that regulate protein degradation as well as its close interrelationship with protein synthesis.",
author = "Fernando Aniento and Enrique Roche and Cuervo, {Ana Maria} and Erwin Knecht",
year = "1993",
month = "5",
day = "15",
language = "English (US)",
volume = "268",
pages = "10463--10470",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "14",

}

TY - JOUR

T1 - Uptake and degradation of glyceraldehyde-3-phosphate dehydrogenase by rat liver lysosomes

AU - Aniento, Fernando

AU - Roche, Enrique

AU - Cuervo, Ana Maria

AU - Knecht, Erwin

PY - 1993/5/15

Y1 - 1993/5/15

N2 - The molecular mechanisms involved in the degradation of individual cellular proteins are probably unique and characteristic. We have investigated in rat liver the degradation of glyceraldehyde-3-phosphate dehydrogenase, an abundant cytosolic enzyme of the glycolytic pathway. Immunoblot analysis of isolated liver lysosomes from rats treated with lysosomal inhibitors show that this protein is degraded, at least in part, by a lysosomal pathway. This pathway was further investigated by incubating the enzyme with lysosomes in a cell-free system, followed by proteolysis measurements, sodium dodecyl sulfate-polyacrylamide gel electrophoresis of lysosomes, and electron microscopic immunocytochemistry. We postulate that the degradative mechanism of glyceraldehyde-3-phosphate dehydrogenase includes a temperature-dependent lysosomal pathway, different from classical nonspecific macroautophagy. The postulated pathway involves: binding of the enzyme to the lysosomal membrane, entry into the lysosomal matrix, and degradation. This cell-free system, which can also incorporate in vitro synthesized proteins, should allow further advances toward clarifying the complex signals that regulate protein degradation as well as its close interrelationship with protein synthesis.

AB - The molecular mechanisms involved in the degradation of individual cellular proteins are probably unique and characteristic. We have investigated in rat liver the degradation of glyceraldehyde-3-phosphate dehydrogenase, an abundant cytosolic enzyme of the glycolytic pathway. Immunoblot analysis of isolated liver lysosomes from rats treated with lysosomal inhibitors show that this protein is degraded, at least in part, by a lysosomal pathway. This pathway was further investigated by incubating the enzyme with lysosomes in a cell-free system, followed by proteolysis measurements, sodium dodecyl sulfate-polyacrylamide gel electrophoresis of lysosomes, and electron microscopic immunocytochemistry. We postulate that the degradative mechanism of glyceraldehyde-3-phosphate dehydrogenase includes a temperature-dependent lysosomal pathway, different from classical nonspecific macroautophagy. The postulated pathway involves: binding of the enzyme to the lysosomal membrane, entry into the lysosomal matrix, and degradation. This cell-free system, which can also incorporate in vitro synthesized proteins, should allow further advances toward clarifying the complex signals that regulate protein degradation as well as its close interrelationship with protein synthesis.

UR - http://www.scopus.com/inward/record.url?scp=0027280820&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0027280820&partnerID=8YFLogxK

M3 - Article

C2 - 8486700

AN - SCOPUS:0027280820

VL - 268

SP - 10463

EP - 10470

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 14

ER -