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PMID: 10639150 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Enzymatic reduction of disulfide bonds in lysosomes: characterization of a gamma-interferon-inducible lysosomal thiol reductase (GILT).

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 97 ·No. 2 ·2000-01-18 ·Pages 745-50

Arunachalam B, Phan UT, Geuze HJ, Cresswell P

Abstract

Proteins internalized into the endocytic pathway are usually degraded. Efficient proteolysis requires denaturation, induced by acidic conditions within lysosomes, and reduction of inter- and intrachain disulfide bonds. Cytosolic reduction is mediated enzymatically by thioredoxin, but the mechanism of lysosomal reduction is unknown. We describe here a lysosomal thiol reductase optimally active at low pH and capable of catalyzing disulfide bond reduction both in vivo and in vitro. The active site, determined by mutagenesis, consists of a pair of cysteine residues separated by two amino acids, similar to other enzymes of the thioredoxin family. The enzyme is a soluble glycoprotein that is synthesized as a precursor. After delivery into the endosomal/lysosomal system by the mannose 6-phosphate receptor, N- and C-terminal prosequences are removed. The enzyme is expressed constitutively in antigen-presenting cells and induced by IFN-gamma in other cell types, suggesting a potentially important role in antigen processing.

MeSH Terms
Amino Acid Sequence Animals Base Sequence Binding Sites/genetics COS Cells DNA, Complementary/chemistry,genetics Disulfides/metabolism Endosomes/enzymology,ultrastructure Enzyme Induction/drug effects Humans Hydrogen-Ion Concentration Interferon-gamma/pharmacology Lysosomes/enzymology Mannosephosphates/metabolism Microscopy, Immunoelectron Molecular Sequence Data Mutagenesis Oxidation-Reduction Protein Disulfide Reductase (Glutathione)/biosynthesis,genetics,metabolism Protein Processing, Post-Translational Sequence Analysis, DNA Tumor Cells, Cultured/drug effects,enzymology,ultrastructure
Chemicals
DNA, Complementary Disulfides Mannosephosphates mannose-6-phosphate Interferon-gamma Protein Disulfide Reductase (Glutathione)
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Arunachalam B
Section of Immunobiology, Howard Hughes Medical Institute, Yale University School of Medicine, P.O. Box 208011, New Haven, CT 06520-8011, USA.
Phan U T
Geuze H J
Cresswell P
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.97.2.745
Published
2000-01-18
Pages
745-50
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC15401
Subset
IM
Grants
NIAID NIH HHS · R01 AI023081 · United States
NIAID NIH HHS · R37 AI023081 · United States
NIAID NIH HHS · AI23081 · United States
Databases
GENBANK
AF097362
External Links
PubMed source DOI: 10.1073/pnas.97.2.745 PMC Full Text View journal details
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