The effect of protein complexation on the mechanical stability of Im9

Eleanore Hann; Nadine Kirkpatrick; Colin Kleanthous; D Alastair Smith; Sheena E Radford; David J Brockwell

doi:10.1529/biophysj.106.102475

The effect of protein complexation on the mechanical stability of Im9

Eleanore Hann, Nadine Kirkpatrick, Colin Kleanthous, D Alastair Smith, Sheena E Radford, David J Brockwell

Biology

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

Abstract

Force mode microscopy can be used to examine the effect of mechanical manipulation on the noncovalent interactions that stabilize proteins and their complexes. Here we describe the effect of complexation by the high affinity protein ligand E9 on the mechanical resistance of the simple four-helical protein, Im9. When concatenated into a construct of alternating I27 domains, Im9 unfolded below the thermal noise limit of the instrument (similar to 20 pN). Complexation of E9 had little effect on the mechanical resistance of Im9 (unfolding force similar to 30 pN) despite the high avidity of this complex (K-d similar to 10 fM).

Original language	English
Pages (from-to)	L79-81
Number of pages	3
Journal	Biophysical Journal
Volume	92
Issue number	9
DOIs	https://doi.org/10.1529/biophysj.106.102475
Publication status	Published - 1 May 2007

Keywords

DIHYDROFOLATE-REDUCTASE
BINDING

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10.1529/biophysj.106.102475

Cite this

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title = "The effect of protein complexation on the mechanical stability of Im9",

abstract = "Force mode microscopy can be used to examine the effect of mechanical manipulation on the noncovalent interactions that stabilize proteins and their complexes. Here we describe the effect of complexation by the high affinity protein ligand E9 on the mechanical resistance of the simple four-helical protein, Im9. When concatenated into a construct of alternating I27 domains, Im9 unfolded below the thermal noise limit of the instrument (similar to 20 pN). Complexation of E9 had little effect on the mechanical resistance of Im9 (unfolding force similar to 30 pN) despite the high avidity of this complex (K-d similar to 10 fM).",

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AU - Hann, Eleanore

AU - Kirkpatrick, Nadine

AU - Kleanthous, Colin

AU - Smith, D Alastair

AU - Radford, Sheena E

AU - Brockwell, David J

PY - 2007/5/1

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AB - Force mode microscopy can be used to examine the effect of mechanical manipulation on the noncovalent interactions that stabilize proteins and their complexes. Here we describe the effect of complexation by the high affinity protein ligand E9 on the mechanical resistance of the simple four-helical protein, Im9. When concatenated into a construct of alternating I27 domains, Im9 unfolded below the thermal noise limit of the instrument (similar to 20 pN). Complexation of E9 had little effect on the mechanical resistance of Im9 (unfolding force similar to 30 pN) despite the high avidity of this complex (K-d similar to 10 fM).

KW - DIHYDROFOLATE-REDUCTASE

KW - BINDING

U2 - 10.1529/biophysj.106.102475

DO - 10.1529/biophysj.106.102475

M3 - Article

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SN - 0006-3495

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SP - L79-81

JO - Biophysical Journal

JF - Biophysical Journal

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