DOI: 10.1083/jcb.137.3.555. Multiple Determinants Direct the Orientation of Signal–Anchor Proteins: The Topogenic Role of the Hydrophobic Signal Domain

Multiple Determinants Direct the Orientation of Signal–Anchor Proteins: The Topogenic Role of the Hydrophobic Signal Domain

10.1083/jcb.137.3.555

Crossref journal-article

Rockefeller University Press

The Journal of Cell Biology (291)

Abstract

The orientation of signal–anchor proteins in the endoplasmic reticulum membrane is largely determined by the charged residues flanking the apolar, membrane-spanning domain and is influenced by the folding properties of the NH2-terminal sequence. However, these features are not generally sufficient to ensure a unique topology. The topogenic role of the hydrophobic signal domain was studied in vivo by expressing mutants of the asialoglycoprotein receptor subunit H1 in COS-7 cells. By replacing the 19-residue transmembrane segment of wild-type and mutant H1 by stretches of 7–25 leucine residues, we found that the length and hydrophobicity of the apolar sequence significantly affected protein orientation. Translocation of the NH2 terminus was favored by long, hydrophobic sequences and translocation of the COOH terminus by short ones. The topogenic contributions of the transmembrane domain, the flanking charges, and a hydrophilic NH2-terminal portion were additive. In combination these determinants were sufficient to achieve unique membrane insertion in either orientation.

Bibliography

Wahlberg, J. M., & Spiess, M. (1997). Multiple Determinants Direct the Orientation of SignalâAnchor Proteins: The Topogenic Role of the Hydrophobic Signal Domain. The Journal of Cell Biology, 137(3), 555â562.

Authors 2

Johanna M. Wahlberg (first)
Martin Spiess (additional)

References 32 Referenced 117

10.1016/S0021-9258(18)42580-X / J Biol Chem / The role of the N-region in signal sequence and signal-anchor function by Andrews (1992)
10.1016/S0021-9258(17)35269-9 / J Biol Chem / Charged residues are major determinants of the transmembrane orientation of a signal-anchor sequence by Beltzer (1991)
10.1016/0076-6879(87)52074-2 / Methods Enzymol / Use of eukaryotic expression technology in the functional analysis of cloned genes by Cullen (1987)
10.1016/0968-0004(90)90047-F / Trends Biochem Sci / Positively charged residues are important determinants of membrane protein topology by Dalbey (1990)
10.1002/j.1460-2075.1995.tb00321.x / EMBO (Eur Mol Biol Organ) J / Transmembrane orientation of signal-anchor proteins is affected by the folding state but not the size of the N-terminal domain by Denzer (1995)
10.1016/0092-8674(86)90786-5 / Cell / Replacement of insulin receptor tyrosine residues 1162 and 1163 compromises insulin-stimulated kinase activity and uptake of 2-deoxyglucose by Ellis (1986)
10.1016/0092-8674(85)90107-2 / Cell / Translocation of secretory proteins across the microsomal membrane occurs through an environment accessible to aqueous perturbants by Gilmore (1985)
10.1073/pnas.86.15.5786 / Proc Natl Acad Sci USA / Predicting the orientation of eukaryotic membrane-spanning proteins by Hartmann (1989)
10.1016/0022-2836(82)90515-0 / J Mol Biol / A simple method for displaying the hydrophobic character of a protein by Kyte (1982)
10.1083/jcb.107.2.457 / J Cell Biol / Signals for the incorporation and orientation of cytochrome P450 in the endoplasmic reticulum membrane by Monier (1988)
10.1002/j.1460-2075.1995.tb00151.x / EMBO (Eur Mol Biol Organ) J / An investigation of the role of transmembrane domains in Golgi protein retention by Munro (1995)
10.1083/jcb.126.5.1127 / J Cell Biol / The COOH-terminal ends of internal signal and signal-anchor sequences are positioned differently in the ER translocase by Nilsson (1994)
10.1074/jbc.271.12.7187 / J Biol Chem / Differential effects of changes in the length of a signal/anchor domain on membrane insertion, subunit assembly, and intracellular transport of a type II integral membrane protein by Parks (1996)
10.1016/0092-8674(91)90507-U / Cell / Topology of eukaryotic type-II membrane proteins—importance of N-terminal positively charged residues flanking the hydrophobic domain by Parks (1991)
10.1016/S0021-9258(17)46740-8 / J Biol Chem / Role of NH2-terminal positively charged residues in establishing membrane protein topology by Parks (1993)
10.1073/pnas.89.1.16 / Proc Natl Acad Sci USA / Functions of signal and signal-anchor sequences are determined by the balance between the hydrophobic segment and the N-terminal charge by Sakaguchi (1992)
10.1002/j.1460-2075.1990.tb07414.x / EMBO (Eur Mol Biol Organ) J / The amino-terminal structures that determine topological orientation of cytochrome-P-450 in microsomal membrane by Sato (1990)
10.1016/S0021-9258(18)37474-X / J Biol Chem / Deletion of the amino-terminal domain of asialoglycoprotein receptor H1 allows cleavage of the internal signal sequence by Schmid (1988)
10.1016/0014-5793(95)00551-J / FEBS (Fed Eur Biochem Soc) Lett / Heads or tails—what determines the orientation of proteins in the membrane by Spiess (1995)
10.1016/0092-8674(86)90496-4 / Cell / An internal signal sequence: the asialoglycoprotein receptor membrane anchor by Spiess (1986)
10.1002/j.1460-2075.1987.tb02560.x / EMBO (Eur Mol Biol Organ) J / Deletion analysis of the internal signalanchor domain of the human asialoglycoprotein receptor H1 by Spiess (1987)
10.1016/S0021-9258(18)89497-2 / J Biol Chem / Sequence of human asialoglycoprotein receptor cDNA. An internal signal sequence for membrane insertion by Spiess (1985)
10.1083/jcb.108.4.1237 / J Cell Biol / NH2-terminal substitutions of basic amino acids induce translocation across the microsomal membrane and glycosylation of rabbit cytochrome P450IIC2 by Szczesna-Skorupa (1989)
10.1073/pnas.85.3.738 / Proc Natl Acad Sci USA / Positive charges at the NH2terminus convert the membrane-anchor signal peptide of cytochrome P-450 to a secretory peptide by Szczesna-Skorupa (1988)
10.1083/jcb.134.1.25 / J Cell Biol / Signal sequence-dependent function of the TRAM protein during early phases of protein transport across the endoplasmic reticulum membrane by Voigt (1996)
10.1002/j.1460-2075.1986.tb04601.x / EMBO (Eur Mol Biol Organ) J / The distribution of positively charged residues in bacterial inner membrane proteins correlates with the trans-membrane topology by von Heijne (1986)
10.1038/341456a0 / Nature (Lond) / Control of topology and mode of assembly of a polytopic membrane protein by positively charged residues by von Heijne (1989)
10.1146/annurev.bb.23.060194.001123 / Annu Rev Biophys Biomol Struc / Membrane proteins: from sequence to structure by von Heijne (1994)
10.1111/j.1432-1033.1988.tb14150.x / Eur J Biochem / Topogenic signals in integral membrane proteins by von Heijne (1988)
10.1093/protein/4.2.109 / Protein Eng / Membrane proteins: from sequence to structure by von Heijne (1990)
10.1083/jcb.130.2.285 / J Cell Biol / transGolgi retention of a plasma membrane protein: mutations in the cytoplasmic domain of the asialoglycoprotein receptor subunit H1 result in trans-Golgi retention by Wahlberg (1995)
10.1146/annurev.cb.10.110194.000511 / Annu Rev Cell Biol / Signal sequence recognition and protein targeting to the endoplasmic reticulum membrane by Walter (1994)

Dates

Type	When
Created	23 years ago (July 26, 2002, 12:47 p.m.)
Deposited	2 years, 1 month ago (July 22, 2023, 2:50 a.m.)
Indexed	2 weeks, 4 days ago (Aug. 5, 2025, 8:36 a.m.)
Issued	28 years, 3 months ago (May 5, 1997)
Published	28 years, 3 months ago (May 5, 1997)
Published Online	28 years, 3 months ago (May 5, 1997)
Published Print	28 years, 3 months ago (May 5, 1997)

Funders 0

None

BibTeX

@article{Wahlberg_1997, title={Multiple Determinants Direct the Orientation of Signal–Anchor Proteins: The Topogenic Role of the Hydrophobic Signal Domain}, volume={137}, ISSN={1540-8140}, url={http://dx.doi.org/10.1083/jcb.137.3.555}, DOI={10.1083/jcb.137.3.555}, number={3}, journal={The Journal of Cell Biology}, publisher={Rockefeller University Press}, author={Wahlberg, Johanna M. and Spiess, Martin}, year={1997}, month=may, pages={555–562} }

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