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IQ motif selectivity in human IQGAP1: binding of myosin essential light chain and S100B

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Abstract

IQGAPs are cytoskeletal scaffolding proteins which link signalling pathways to the reorganisation of actin and microtubules. Human IQGAP1 has four IQ motifs each of which binds to calmodulin. The same region has been implicated in binding to two calmodulin-like proteins, the myosin essential light chain Mlc1sa and the calcium and zinc ion binding protein S100B. Using synthetic peptides corresponding to the four IQ motifs of human IQGAP1, we showed by native gel electrophoresis that only the first IQ motif interacts with Mlc1sa. This IQ motif, and also the fourth, interacts with the budding yeast myosin essential light chain Mlc1p. The first and second IQ motifs interact with S100B in the presence of calcium ions. This clearly establishes that S100B can interact with its targets through IQ motifs in addition to interacting via previously reported sequences. These results are discussed in terms of the function of IQGAP1 and IQ motif recognition.

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Abbreviations

APC:

Adenomatous polyposis coli

DIEA:

N,N-diisopropylethylamine

FhCaM2:

Fasciola hepatica type 2 calmodulin

Fmoc:

9-Fluorenylmethoxycarbonyl

HATU:

2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyl uronium hexafluorophosphate methanaminium

IQ1, IQ2, IQ3, IQ4:

The first, second, third and fourth IQ motifs (as numbered from the N-terminus), respectively, of human IQGAP1

K d,app :

The apparent disassociation constant under the conditions of native gel electrophoresis

LB:

Luria-Bertani

TFA:

Trifluoroacetic acid

VEGFR2:

Vascular endothelial growth factor receptor

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Acknowledgements

We thank Sean Russell for providing FhCaM2 protein which was used as a control in these experiments and Maelíosa Mc Crudden for preliminary work on the purification of Mlc1sa. This work was funded in part by a grant from the BBSRC (reference BB/D000394/1). Sevvel Pathmanathan gratefully acknowledges a studentship from Queen’s University, Belfast and the University of Jaffna (Sri Lanka) for giving him leave of absence to complete this work.

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  1. Medical Biology Centre, School of Biological Sciences, Queen’s University Belfast, 97 Lisburn Road, Belfast, BT9 7BL, UK

    Sevvel Pathmanathan, Sarah F. Elliott, Sara McSwiggen, Brett Greer, Pat Harriott & David J. Timson

  2. School of Medicine & Dentistry, Queen’s University Belfast, Whitla Medical Building, 97 Lisburn Road, Belfast, BT9 7BL, UK

    G. Brent Irvine

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  1. Sevvel Pathmanathan
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Correspondence to David J. Timson.

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Pathmanathan, S., Elliott, S.F., McSwiggen, S. et al. IQ motif selectivity in human IQGAP1: binding of myosin essential light chain and S100B. Mol Cell Biochem 318, 43–51 (2008). https://doi.org/10.1007/s11010-008-9855-9

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