Dual role of the cysteine-string domain in membrane binding and palmitoylation-dependent sorting of the molecular chaperone cysteine-string protein

Jennifer Greaves, Luke H Chamberlain

Research output: Contribution to journalArticle

42 Citations (Scopus)

Abstract

S-palmitoylation occurs on intracellular membranes and, therefore, membrane anchoring of proteins must precede palmitate transfer. However, a number of palmitoylated proteins lack any obvious membrane targeting motifs and it is unclear how this class of proteins become membrane associated before palmitoylation. Cysteine-string protein (CSP), which is extensively palmitoylated on a "string" of 14 cysteine residues, is an example of such a protein. In this study, we have investigated the mechanisms that govern initial membrane targeting, palmitoylation, and membrane trafficking of CSP. We identified a hydrophobic 31 amino acid domain, which includes the cysteine-string, as a membrane-targeting motif that associates predominantly with endoplasmic reticulum (ER) membranes. Cysteine residues in this domain are not merely sites for the addition of palmitate groups, but play an essential role in membrane recognition before palmitoylation. Membrane association of the cysteine-string domain is not sufficient to trigger palmitoylation, which requires additional downstream residues that may regulate the membrane orientation of the cysteine-string domain. CSP palmitoylation-deficient mutants remain "trapped" in the ER, suggesting that palmitoylation may regulate ER exit and correct intracellular sorting of CSP. These results reveal a dual function of the cysteine-string domain: initial membrane binding and palmitoylation-dependent sorting.

Original languageEnglish
Pages (from-to)4748-4759
Number of pages12
JournalMolecular Biology of the Cell
Volume17
Issue number11
Early online date30 Aug 2006
DOIs
Publication statusPublished - 1 Nov 2006
Externally publishedYes

Fingerprint

Lipoylation
Molecular Chaperones
Cysteine
Membranes
Endoplasmic Reticulum
Palmitates
Membrane Proteins
cysteine string protein
Intracellular Membranes
Proteins
Amino Acids

Keywords

  • Amino Acid Sequence
  • Animals
  • Biomarkers
  • Cell Membrane
  • Cysteine
  • Endoplasmic Reticulum
  • Green Fluorescent Proteins
  • HSP40 Heat-Shock Proteins
  • Membrane Proteins
  • Molecular Chaperones
  • Molecular Sequence Data
  • Mutant Proteins
  • PC12 Cells
  • Palmitates
  • Protein Binding
  • Protein Structure, Tertiary
  • Protein Transport
  • Rats
  • Recombinant Fusion Proteins
  • Sequence Analysis, Protein
  • Structure-Activity Relationship
  • Journal Article
  • Research Support, Non-U.S. Gov't

Cite this

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title = "Dual role of the cysteine-string domain in membrane binding and palmitoylation-dependent sorting of the molecular chaperone cysteine-string protein",
abstract = "S-palmitoylation occurs on intracellular membranes and, therefore, membrane anchoring of proteins must precede palmitate transfer. However, a number of palmitoylated proteins lack any obvious membrane targeting motifs and it is unclear how this class of proteins become membrane associated before palmitoylation. Cysteine-string protein (CSP), which is extensively palmitoylated on a {"}string{"} of 14 cysteine residues, is an example of such a protein. In this study, we have investigated the mechanisms that govern initial membrane targeting, palmitoylation, and membrane trafficking of CSP. We identified a hydrophobic 31 amino acid domain, which includes the cysteine-string, as a membrane-targeting motif that associates predominantly with endoplasmic reticulum (ER) membranes. Cysteine residues in this domain are not merely sites for the addition of palmitate groups, but play an essential role in membrane recognition before palmitoylation. Membrane association of the cysteine-string domain is not sufficient to trigger palmitoylation, which requires additional downstream residues that may regulate the membrane orientation of the cysteine-string domain. CSP palmitoylation-deficient mutants remain {"}trapped{"} in the ER, suggesting that palmitoylation may regulate ER exit and correct intracellular sorting of CSP. These results reveal a dual function of the cysteine-string domain: initial membrane binding and palmitoylation-dependent sorting.",
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TY - JOUR

T1 - Dual role of the cysteine-string domain in membrane binding and palmitoylation-dependent sorting of the molecular chaperone cysteine-string protein

AU - Greaves, Jennifer

AU - Chamberlain, Luke H

PY - 2006/11/1

Y1 - 2006/11/1

N2 - S-palmitoylation occurs on intracellular membranes and, therefore, membrane anchoring of proteins must precede palmitate transfer. However, a number of palmitoylated proteins lack any obvious membrane targeting motifs and it is unclear how this class of proteins become membrane associated before palmitoylation. Cysteine-string protein (CSP), which is extensively palmitoylated on a "string" of 14 cysteine residues, is an example of such a protein. In this study, we have investigated the mechanisms that govern initial membrane targeting, palmitoylation, and membrane trafficking of CSP. We identified a hydrophobic 31 amino acid domain, which includes the cysteine-string, as a membrane-targeting motif that associates predominantly with endoplasmic reticulum (ER) membranes. Cysteine residues in this domain are not merely sites for the addition of palmitate groups, but play an essential role in membrane recognition before palmitoylation. Membrane association of the cysteine-string domain is not sufficient to trigger palmitoylation, which requires additional downstream residues that may regulate the membrane orientation of the cysteine-string domain. CSP palmitoylation-deficient mutants remain "trapped" in the ER, suggesting that palmitoylation may regulate ER exit and correct intracellular sorting of CSP. These results reveal a dual function of the cysteine-string domain: initial membrane binding and palmitoylation-dependent sorting.

AB - S-palmitoylation occurs on intracellular membranes and, therefore, membrane anchoring of proteins must precede palmitate transfer. However, a number of palmitoylated proteins lack any obvious membrane targeting motifs and it is unclear how this class of proteins become membrane associated before palmitoylation. Cysteine-string protein (CSP), which is extensively palmitoylated on a "string" of 14 cysteine residues, is an example of such a protein. In this study, we have investigated the mechanisms that govern initial membrane targeting, palmitoylation, and membrane trafficking of CSP. We identified a hydrophobic 31 amino acid domain, which includes the cysteine-string, as a membrane-targeting motif that associates predominantly with endoplasmic reticulum (ER) membranes. Cysteine residues in this domain are not merely sites for the addition of palmitate groups, but play an essential role in membrane recognition before palmitoylation. Membrane association of the cysteine-string domain is not sufficient to trigger palmitoylation, which requires additional downstream residues that may regulate the membrane orientation of the cysteine-string domain. CSP palmitoylation-deficient mutants remain "trapped" in the ER, suggesting that palmitoylation may regulate ER exit and correct intracellular sorting of CSP. These results reveal a dual function of the cysteine-string domain: initial membrane binding and palmitoylation-dependent sorting.

KW - Amino Acid Sequence

KW - Animals

KW - Biomarkers

KW - Cell Membrane

KW - Cysteine

KW - Endoplasmic Reticulum

KW - Green Fluorescent Proteins

KW - HSP40 Heat-Shock Proteins

KW - Membrane Proteins

KW - Molecular Chaperones

KW - Molecular Sequence Data

KW - Mutant Proteins

KW - PC12 Cells

KW - Palmitates

KW - Protein Binding

KW - Protein Structure, Tertiary

KW - Protein Transport

KW - Rats

KW - Recombinant Fusion Proteins

KW - Sequence Analysis, Protein

KW - Structure-Activity Relationship

KW - Journal Article

KW - Research Support, Non-U.S. Gov't

U2 - 10.1091/mbc.e06-03-0183

DO - 10.1091/mbc.e06-03-0183

M3 - Article

VL - 17

SP - 4748

EP - 4759

JO - The Journal of biophysical and biochemical cytology

JF - The Journal of biophysical and biochemical cytology

SN - 1465-7392

IS - 11

ER -