A substrate-optimized electrophoretic mobility shift assay for ADAM12

Alexander Kotzsch; Tine Skovgaard; Uwe Buus; Simon Andersen; Kanchan Devkota; Jens Berthelsen

doi:10.1016/j.ab.2014.02.008

A substrate-optimized electrophoretic mobility shift assay for ADAM12

Alexander Kotzsch, Tine Skovgaard, Uwe Buus, Simon Andersen, Kanchan Devkota, Jens Berthelsen

Abstract

ADAM12 belongs to the A disintegrin and metalloprotease (ADAM) family of secreted sheddases activating extracellular growth factors such as epidermal growth factor receptor (EGFR) ligands and tumor necrosis factor-alpha (TNF-α). ADAM proteases, most notably ADAM17 (TNF-α-converting enzyme), have long been investigated as pharmaceutical drug targets; however, due to lack of potency and in vivo side effects, none of the small-molecule inhibitors discovered so far has made it beyond clinical testing. Ongoing research on novel selective inhibitors of ADAMs requires reliable biochemical assays to validate molecular probes from large-scale screening efforts. Here we describe an electrophoretic mobility shift assay for ADAM12 based on the identification of an optimized peptide substrate that is characterized by excellent performance and reproducibility.

Originalsprog	Engelsk
Tidsskrift	Analytical Biochemistry
Vol/bind	452
Sider (fra-til)	34-42
Antal sider	9
ISSN	0003-2697
DOI	https://doi.org/10.1016/j.ab.2014.02.008
Status	Udgivet - 1 maj 2014

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10.1016/j.ab.2014.02.008

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title = "A substrate-optimized electrophoretic mobility shift assay for ADAM12",

abstract = "ADAM12 belongs to the A disintegrin and metalloprotease (ADAM) family of secreted sheddases activating extracellular growth factors such as epidermal growth factor receptor (EGFR) ligands and tumor necrosis factor-alpha (TNF-α). ADAM proteases, most notably ADAM17 (TNF-α-converting enzyme), have long been investigated as pharmaceutical drug targets; however, due to lack of potency and in vivo side effects, none of the small-molecule inhibitors discovered so far has made it beyond clinical testing. Ongoing research on novel selective inhibitors of ADAMs requires reliable biochemical assays to validate molecular probes from large-scale screening efforts. Here we describe an electrophoretic mobility shift assay for ADAM12 based on the identification of an optimized peptide substrate that is characterized by excellent performance and reproducibility.",

author = "Alexander Kotzsch and Tine Skovgaard and Uwe Buus and Simon Andersen and Kanchan Devkota and Jens Berthelsen",

year = "2014",

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doi = "10.1016/j.ab.2014.02.008",

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T1 - A substrate-optimized electrophoretic mobility shift assay for ADAM12

AU - Kotzsch, Alexander

AU - Skovgaard, Tine

AU - Buus, Uwe

AU - Andersen, Simon

AU - Devkota, Kanchan

AU - Berthelsen, Jens

PY - 2014/5/1

Y1 - 2014/5/1

N2 - ADAM12 belongs to the A disintegrin and metalloprotease (ADAM) family of secreted sheddases activating extracellular growth factors such as epidermal growth factor receptor (EGFR) ligands and tumor necrosis factor-alpha (TNF-α). ADAM proteases, most notably ADAM17 (TNF-α-converting enzyme), have long been investigated as pharmaceutical drug targets; however, due to lack of potency and in vivo side effects, none of the small-molecule inhibitors discovered so far has made it beyond clinical testing. Ongoing research on novel selective inhibitors of ADAMs requires reliable biochemical assays to validate molecular probes from large-scale screening efforts. Here we describe an electrophoretic mobility shift assay for ADAM12 based on the identification of an optimized peptide substrate that is characterized by excellent performance and reproducibility.

AB - ADAM12 belongs to the A disintegrin and metalloprotease (ADAM) family of secreted sheddases activating extracellular growth factors such as epidermal growth factor receptor (EGFR) ligands and tumor necrosis factor-alpha (TNF-α). ADAM proteases, most notably ADAM17 (TNF-α-converting enzyme), have long been investigated as pharmaceutical drug targets; however, due to lack of potency and in vivo side effects, none of the small-molecule inhibitors discovered so far has made it beyond clinical testing. Ongoing research on novel selective inhibitors of ADAMs requires reliable biochemical assays to validate molecular probes from large-scale screening efforts. Here we describe an electrophoretic mobility shift assay for ADAM12 based on the identification of an optimized peptide substrate that is characterized by excellent performance and reproducibility.

U2 - 10.1016/j.ab.2014.02.008

DO - 10.1016/j.ab.2014.02.008

M3 - Journal article

C2 - 24534253

SN - 0003-2697

VL - 452

SP - 34

EP - 42

JO - Analytical Biochemistry

JF - Analytical Biochemistry

ER -

A substrate-optimized electrophoretic mobility shift assay for ADAM12

Abstract

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