Universität zu Köln

Moss, Marcia L., Minond, Dmitriy ORCID: 0000-0003-2473-5944, Yoneyama, Toshie, Hansen, Hinrich P., Vujanovic, Nikola and Rasmussen, Fred H. (2016). An improved fluorescent substrate for assaying soluble and membrane-associated ADAM family member activities. Anal. Biochem., 507. S. 13 - 18. SAN DIEGO: ACADEMIC PRESS INC ELSEVIER SCIENCE. ISSN 1096-0309

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Abstract

A fluorescent resonance energy transfer substrate with improved sensitivity for ADAM17, -10, and -9 (where ADAM represents a disintegrin and metalloproteinase) has been designed. The new substrate, Dabcyl-Pro-Arg-Ala-Ala-Ala-Homophe-Thr-Ser-Pro-Lys(FAM)-NH2, has specificity constants of 6.3 (+/- 0.3) x 10(4) M-1 s(-1) and 2.4 (+/- 0.3) x 10(3) M-1 s(-1) for ADAM17 and ADAM10, respectively. The substrate is more sensitive than widely used peptides based on the precursor tumor necrosis factor-alpha (TNF-alpha) cleavage site, PEPDAB010 or Dabcyl-Ser-Pro-Leu-Ala-Gln-Ala-Val-Arg-Ser-Ser-Lys(FAM)-NH2 and Mca-Pro-Leu-Ala-Gln-Ala-Val-Dpa-Arg-Ser-Ser-Arg-NH2. ADAM9 also processes the new peptide more than 18-fold better than the TNF-alpha-based substrates. The new substrate has a unique selectivity profile because it is processed less efficiently by ADAMS and MMP1, -2, -3, -8, -9, -12, and -14. This substrate provides a unique tool in which to assess ADAM17, -10, and -9 activities. (C) 2016 Elsevier Inc. All rights reserved.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Moss, Marcia L. UNSPECIFIED UNSPECIFIED UNSPECIFIED Minond, Dmitriy UNSPECIFIED orcid.org/0000-0003-2473-5944 UNSPECIFIED Yoneyama, Toshie UNSPECIFIED UNSPECIFIED UNSPECIFIED Hansen, Hinrich P. UNSPECIFIED UNSPECIFIED UNSPECIFIED Vujanovic, Nikola UNSPECIFIED UNSPECIFIED UNSPECIFIED Rasmussen, Fred H. UNSPECIFIED UNSPECIFIED UNSPECIFIED
URN:	urn:nbn:de:hbz:38-266580
DOI:	10.1016/j.ab.2016.05.001
Journal or Publication Title:	Anal. Biochem.
Volume:	507
Page Range:	S. 13 - 18
Date:	2016
Publisher:	ACADEMIC PRESS INC ELSEVIER SCIENCE
Place of Publication:	SAN DIEGO
ISSN:	1096-0309
Language:	English
Faculty:	Unspecified
Divisions:	Unspecified
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language CRYSTAL-STRUCTURES; INHIBITORS; METALLOPROTEINASES; CANCER Multiple languages Biochemical Research Methods; Biochemistry & Molecular Biology; Chemistry, Analytical Multiple languages
Refereed:	Yes
URI:	http://kups.ub.uni-koeln.de/id/eprint/26658