Membrane insertion of the FYVE domain is modulated by pH

Ju He, Mohsin Vora, Rachel M. Haney, Grigory S. Filonov, Catherine A. Musselman, Christopher G. Burd, Andrei G. Kutateladze, Vladislav Verkhusha, Robert V. Stahelin, Tatiana G. Kutateladze

Research output: Contribution to journalArticle

33 Citations (Scopus)

Abstract

The FYVE domain associates with phosphatidylinositol 3-phosphate [PtdIns(3)P] in membranes of early endosomes and penetrates bilayers. Here, we detail principles of membrane anchoring and show that the FYVE domain insertion into PtdIns(3)P-enriched membranes and membrane-mimetics is substantially increased in acidic conditions. The EEA1 FYVE domain binds to POPC/POPE/ PtdIns(3)P vesicles with a Kd of 49 nM at pH 6.0, however associates ∼24 fold weaker at pH 8.0. The decrease in the affinity is primarily due to much faster dissociation of the protein from the bilayers in basic media. Lowering the pH enhances the interaction of the Hrs, RUFY1, Vps27p and WDFY1 FYVE domains with PtdIns(3)P-containing membranes in vitro and in vivo, indicating that pH-dependency is a general function of the FYVE finger family. The PtdIns(3)P binding and membrane insertion of the FYVE domain is modulated by the two adjacent His residues of the R(R/K)HHCRXCG signature motif. Mutation of either His residue abolishes the pH-sensitivity. Both protonation of the His residues and nonspecific electrostatic contacts stabilize the FYVE domain in the lipid-bound form, promoting its penetration and increasing the membrane residence time.

Original languageEnglish (US)
Pages (from-to)852-860
Number of pages9
JournalProteins: Structure, Function and Bioinformatics
Volume76
Issue number4
DOIs
StatePublished - Sep 2009

Fingerprint

Membranes
Endosomes
Protonation
Static Electricity
Fingers
Electrostatics
phosphatidylinositol 3-phosphate
Lipids
Mutation
Proteins

Keywords

  • FYVE
  • Mechanism
  • Membrane
  • pH dependence
  • Phosphoinositide

ASJC Scopus subject areas

  • Biochemistry
  • Structural Biology
  • Molecular Biology

Cite this

He, J., Vora, M., Haney, R. M., Filonov, G. S., Musselman, C. A., Burd, C. G., ... Kutateladze, T. G. (2009). Membrane insertion of the FYVE domain is modulated by pH. Proteins: Structure, Function and Bioinformatics, 76(4), 852-860. https://doi.org/10.1002/prot.22392

Membrane insertion of the FYVE domain is modulated by pH. / He, Ju; Vora, Mohsin; Haney, Rachel M.; Filonov, Grigory S.; Musselman, Catherine A.; Burd, Christopher G.; Kutateladze, Andrei G.; Verkhusha, Vladislav; Stahelin, Robert V.; Kutateladze, Tatiana G.

In: Proteins: Structure, Function and Bioinformatics, Vol. 76, No. 4, 09.2009, p. 852-860.

Research output: Contribution to journalArticle

He, J, Vora, M, Haney, RM, Filonov, GS, Musselman, CA, Burd, CG, Kutateladze, AG, Verkhusha, V, Stahelin, RV & Kutateladze, TG 2009, 'Membrane insertion of the FYVE domain is modulated by pH', Proteins: Structure, Function and Bioinformatics, vol. 76, no. 4, pp. 852-860. https://doi.org/10.1002/prot.22392
He J, Vora M, Haney RM, Filonov GS, Musselman CA, Burd CG et al. Membrane insertion of the FYVE domain is modulated by pH. Proteins: Structure, Function and Bioinformatics. 2009 Sep;76(4):852-860. https://doi.org/10.1002/prot.22392
He, Ju ; Vora, Mohsin ; Haney, Rachel M. ; Filonov, Grigory S. ; Musselman, Catherine A. ; Burd, Christopher G. ; Kutateladze, Andrei G. ; Verkhusha, Vladislav ; Stahelin, Robert V. ; Kutateladze, Tatiana G. / Membrane insertion of the FYVE domain is modulated by pH. In: Proteins: Structure, Function and Bioinformatics. 2009 ; Vol. 76, No. 4. pp. 852-860.
@article{5a851de7402645fdbd154fdd90ca0725,
title = "Membrane insertion of the FYVE domain is modulated by pH",
abstract = "The FYVE domain associates with phosphatidylinositol 3-phosphate [PtdIns(3)P] in membranes of early endosomes and penetrates bilayers. Here, we detail principles of membrane anchoring and show that the FYVE domain insertion into PtdIns(3)P-enriched membranes and membrane-mimetics is substantially increased in acidic conditions. The EEA1 FYVE domain binds to POPC/POPE/ PtdIns(3)P vesicles with a Kd of 49 nM at pH 6.0, however associates ∼24 fold weaker at pH 8.0. The decrease in the affinity is primarily due to much faster dissociation of the protein from the bilayers in basic media. Lowering the pH enhances the interaction of the Hrs, RUFY1, Vps27p and WDFY1 FYVE domains with PtdIns(3)P-containing membranes in vitro and in vivo, indicating that pH-dependency is a general function of the FYVE finger family. The PtdIns(3)P binding and membrane insertion of the FYVE domain is modulated by the two adjacent His residues of the R(R/K)HHCRXCG signature motif. Mutation of either His residue abolishes the pH-sensitivity. Both protonation of the His residues and nonspecific electrostatic contacts stabilize the FYVE domain in the lipid-bound form, promoting its penetration and increasing the membrane residence time.",
keywords = "FYVE, Mechanism, Membrane, pH dependence, Phosphoinositide",
author = "Ju He and Mohsin Vora and Haney, {Rachel M.} and Filonov, {Grigory S.} and Musselman, {Catherine A.} and Burd, {Christopher G.} and Kutateladze, {Andrei G.} and Vladislav Verkhusha and Stahelin, {Robert V.} and Kutateladze, {Tatiana G.}",
year = "2009",
month = "9",
doi = "10.1002/prot.22392",
language = "English (US)",
volume = "76",
pages = "852--860",
journal = "Proteins: Structure, Function and Bioinformatics",
issn = "0887-3585",
publisher = "Wiley-Liss Inc.",
number = "4",

}

TY - JOUR

T1 - Membrane insertion of the FYVE domain is modulated by pH

AU - He, Ju

AU - Vora, Mohsin

AU - Haney, Rachel M.

AU - Filonov, Grigory S.

AU - Musselman, Catherine A.

AU - Burd, Christopher G.

AU - Kutateladze, Andrei G.

AU - Verkhusha, Vladislav

AU - Stahelin, Robert V.

AU - Kutateladze, Tatiana G.

PY - 2009/9

Y1 - 2009/9

N2 - The FYVE domain associates with phosphatidylinositol 3-phosphate [PtdIns(3)P] in membranes of early endosomes and penetrates bilayers. Here, we detail principles of membrane anchoring and show that the FYVE domain insertion into PtdIns(3)P-enriched membranes and membrane-mimetics is substantially increased in acidic conditions. The EEA1 FYVE domain binds to POPC/POPE/ PtdIns(3)P vesicles with a Kd of 49 nM at pH 6.0, however associates ∼24 fold weaker at pH 8.0. The decrease in the affinity is primarily due to much faster dissociation of the protein from the bilayers in basic media. Lowering the pH enhances the interaction of the Hrs, RUFY1, Vps27p and WDFY1 FYVE domains with PtdIns(3)P-containing membranes in vitro and in vivo, indicating that pH-dependency is a general function of the FYVE finger family. The PtdIns(3)P binding and membrane insertion of the FYVE domain is modulated by the two adjacent His residues of the R(R/K)HHCRXCG signature motif. Mutation of either His residue abolishes the pH-sensitivity. Both protonation of the His residues and nonspecific electrostatic contacts stabilize the FYVE domain in the lipid-bound form, promoting its penetration and increasing the membrane residence time.

AB - The FYVE domain associates with phosphatidylinositol 3-phosphate [PtdIns(3)P] in membranes of early endosomes and penetrates bilayers. Here, we detail principles of membrane anchoring and show that the FYVE domain insertion into PtdIns(3)P-enriched membranes and membrane-mimetics is substantially increased in acidic conditions. The EEA1 FYVE domain binds to POPC/POPE/ PtdIns(3)P vesicles with a Kd of 49 nM at pH 6.0, however associates ∼24 fold weaker at pH 8.0. The decrease in the affinity is primarily due to much faster dissociation of the protein from the bilayers in basic media. Lowering the pH enhances the interaction of the Hrs, RUFY1, Vps27p and WDFY1 FYVE domains with PtdIns(3)P-containing membranes in vitro and in vivo, indicating that pH-dependency is a general function of the FYVE finger family. The PtdIns(3)P binding and membrane insertion of the FYVE domain is modulated by the two adjacent His residues of the R(R/K)HHCRXCG signature motif. Mutation of either His residue abolishes the pH-sensitivity. Both protonation of the His residues and nonspecific electrostatic contacts stabilize the FYVE domain in the lipid-bound form, promoting its penetration and increasing the membrane residence time.

KW - FYVE

KW - Mechanism

KW - Membrane

KW - pH dependence

KW - Phosphoinositide

UR - http://www.scopus.com/inward/record.url?scp=68149182252&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=68149182252&partnerID=8YFLogxK

U2 - 10.1002/prot.22392

DO - 10.1002/prot.22392

M3 - Article

C2 - 19296456

AN - SCOPUS:68149182252

VL - 76

SP - 852

EP - 860

JO - Proteins: Structure, Function and Bioinformatics

JF - Proteins: Structure, Function and Bioinformatics

SN - 0887-3585

IS - 4

ER -