Characterisation of the Rab binding properties of Rab coupling protein (RCP) by site-directed mutagenesis

Andrew J. Lindsay; Mary W. McCaffrey

doi:10.1016/j.febslet.2004.06.068

Characterisation of the Rab binding properties of Rab coupling protein (RCP) by site-directed mutagenesis

School of Biochemistry and Cell Biology

Research output: Contribution to journal › Article › peer-review

Abstract

Rab coupling protein (RCP) is a member of the Rab11-family of interacting proteins (Rab11-FIPs). Family members are characterised by their ability to interact with Rab11. This property is mediated by a conserved Rab binding domain (RBD) located at their carboxy-termini. Several Rab11-FIPs can also interact with other small GTPases. RCP interacts with Rab4 in addition to Rab11. To dissect out the individual properties of the Rab4 and Rab11 interactions with RCP, conserved amino acids within the RBD of RCP were mutated by site-directed mutagenesis. The effect of these mutations on Rab4 and Rab11 binding, and the intracellular localisation of RCP, was examined. Our results indicate that Rab11, rather than Rab4, mediates the intracellular localisation of RCP, and that the class I Rab11-FIPs compete for binding to Rab11.

Original language	English
Pages (from-to)	86-92
Number of pages	7
Journal	FEBS Letters
Volume	571
Issue number	1-3
DOIs	https://doi.org/10.1016/j.febslet.2004.06.068
Publication status	Published - 30 Jul 2004

Keywords

EGF, epidermal growth factor
Endocytic recycling compartment
ERC, endocytic recycling compartment
PMA, phorbol 12-myristate 13-acetate
Rab11-FIP
Rab11-FIP, Rab11-family of interacting proteins
RCP
RCP, Rab coupling protein
SE, sorting endosome

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10.1016/j.febslet.2004.06.068

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@article{c5fa61e6927747fbae12b47ee395cc7f,

title = "Characterisation of the Rab binding properties of Rab coupling protein (RCP) by site-directed mutagenesis",

abstract = "Rab coupling protein (RCP) is a member of the Rab11-family of interacting proteins (Rab11-FIPs). Family members are characterised by their ability to interact with Rab11. This property is mediated by a conserved Rab binding domain (RBD) located at their carboxy-termini. Several Rab11-FIPs can also interact with other small GTPases. RCP interacts with Rab4 in addition to Rab11. To dissect out the individual properties of the Rab4 and Rab11 interactions with RCP, conserved amino acids within the RBD of RCP were mutated by site-directed mutagenesis. The effect of these mutations on Rab4 and Rab11 binding, and the intracellular localisation of RCP, was examined. Our results indicate that Rab11, rather than Rab4, mediates the intracellular localisation of RCP, and that the class I Rab11-FIPs compete for binding to Rab11.",

keywords = "EGF, epidermal growth factor, Endocytic recycling compartment, ERC, endocytic recycling compartment, PMA, phorbol 12-myristate 13-acetate, Rab11-FIP, Rab11-FIP, Rab11-family of interacting proteins, RCP, RCP, Rab coupling protein, SE, sorting endosome",

author = "Lindsay, \{Andrew J.\} and McCaffrey, \{Mary W.\}",

year = "2004",

month = jul,

day = "30",

doi = "10.1016/j.febslet.2004.06.068",

language = "English",

volume = "571",

pages = "86--92",

journal = "FEBS Letters",

issn = "0014-5793",

publisher = "John Wiley and Sons Inc",

number = "1-3",

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TY - JOUR

T1 - Characterisation of the Rab binding properties of Rab coupling protein (RCP) by site-directed mutagenesis

AU - Lindsay, Andrew J.

AU - McCaffrey, Mary W.

PY - 2004/7/30

Y1 - 2004/7/30

N2 - Rab coupling protein (RCP) is a member of the Rab11-family of interacting proteins (Rab11-FIPs). Family members are characterised by their ability to interact with Rab11. This property is mediated by a conserved Rab binding domain (RBD) located at their carboxy-termini. Several Rab11-FIPs can also interact with other small GTPases. RCP interacts with Rab4 in addition to Rab11. To dissect out the individual properties of the Rab4 and Rab11 interactions with RCP, conserved amino acids within the RBD of RCP were mutated by site-directed mutagenesis. The effect of these mutations on Rab4 and Rab11 binding, and the intracellular localisation of RCP, was examined. Our results indicate that Rab11, rather than Rab4, mediates the intracellular localisation of RCP, and that the class I Rab11-FIPs compete for binding to Rab11.

AB - Rab coupling protein (RCP) is a member of the Rab11-family of interacting proteins (Rab11-FIPs). Family members are characterised by their ability to interact with Rab11. This property is mediated by a conserved Rab binding domain (RBD) located at their carboxy-termini. Several Rab11-FIPs can also interact with other small GTPases. RCP interacts with Rab4 in addition to Rab11. To dissect out the individual properties of the Rab4 and Rab11 interactions with RCP, conserved amino acids within the RBD of RCP were mutated by site-directed mutagenesis. The effect of these mutations on Rab4 and Rab11 binding, and the intracellular localisation of RCP, was examined. Our results indicate that Rab11, rather than Rab4, mediates the intracellular localisation of RCP, and that the class I Rab11-FIPs compete for binding to Rab11.

KW - EGF, epidermal growth factor

KW - Endocytic recycling compartment

KW - ERC, endocytic recycling compartment

KW - PMA, phorbol 12-myristate 13-acetate

KW - Rab11-FIP

KW - Rab11-FIP, Rab11-family of interacting proteins

KW - RCP

KW - RCP, Rab coupling protein

KW - SE, sorting endosome

UR - https://www.scopus.com/pages/publications/3342887805

U2 - 10.1016/j.febslet.2004.06.068

DO - 10.1016/j.febslet.2004.06.068

M3 - Article

C2 - 15280022

AN - SCOPUS:3342887805

SN - 0014-5793

VL - 571

SP - 86

EP - 92

JO - FEBS Letters

JF - FEBS Letters

IS - 1-3

ER -

Characterisation of the Rab binding properties of Rab coupling protein (RCP) by site-directed mutagenesis

Abstract

Keywords

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