Dihydroisoxazole inhibitors of Anopheles gambiae seminal transglutaminase AgTG3

Malaria Journal

Research

Cornelius Klöck¹  Chaitan Khosla¹  Binh V Le²  Alexandra Schatz²  Richard HG Baxter³  Evdoxia G Kakani⁴  Flaminia Catteruccia⁴  Jennifer B Nguyen⁵ 

[1] Department of Chemistry, Stanford University, 94305, Stanford, CA, USA;Department of Chemistry, Yale University, P.O. Box 208107, 06520-8107, New Haven, CT, USA;Department of Chemistry, Yale University, P.O. Box 208107, 06520-8107, New Haven, CT, USA;Department of Molecular Biophysics and Biochemistry, Yale University, 06511, New Haven, CT, USA;Department of Immunology and Infectious Diseases, Harvard School of Public Health, 02115, Boston, MA, USA;Dipartimento di Medicina Sperimentale e Scienze Biochimiche, Università degli Studi di Perugia, 06123, Perugia, Italy;Department of Molecular Biophysics and Biochemistry, Yale University, 06511, New Haven, CT, USA;

关键词: Anopheles Gambiae;    Tobacco Etch Virus;    Human African Trypanosomiasis;    Sterile Insect Technique;    Active Site Cysteine;   

DOI  :  10.1186/1475-2875-13-210

 received in 2014-02-13, accepted in 2014-05-23,  发布年份 2014

来源: Springer

【 摘 要 】

BackgroundCurrent vector-based malaria control strategies are threatened by the rise of biochemical and behavioural resistance in mosquitoes. Researching mosquito traits of immunity and fertility is required to find potential targets for new vector control strategies. The seminal transglutaminase AgTG3 coagulates male Anopheles gambiae seminal fluids, forming a ‘mating plug’ that is required for male reproductive success. Inhibitors of AgTG3 can be useful both as chemical probes of A. gambiae reproductive biology and may further the development of new chemosterilants for mosquito population control.MethodsA targeted library of 3-bromo-4,5-dihydroxoisoxazole inhibitors were synthesized and screened for inhibition of AgTG3 in a fluorescent, plate-based assay. Positive hits were tested for in vitro activity using cross-linking and mass spectrometry, and in vivo efficacy in laboratory mating assays.ResultsA targeted chemical library was screened for inhibition of AgTG3 in a fluorescent plate-based assay using its native substrate, plugin. Several inhibitors were identified with IC50 < 10 μM. Preliminary structure-activity relationships within the library support the stereo-specificity and preference for aromatic substituents in the chemical scaffold. Both inhibition of plugin cross-linking and covalent modification of the active site cysteine of AgTG3 were verified. Administration of an AgTG3 inhibitor to A. gambiae males by intrathoracic injection led to a 15% reduction in mating plug transfer in laboratory mating assays.ConclusionsA targeted screen has identified chemical inhibitors of A. gambiae transglutaminase 3 (AgTG3). The most potent inhibitors are known inhibitors of human transglutaminase 2, suggesting a common binding pose may exist within the active site of both enzymes. Future efforts to develop additional inhibitors will provide chemical tools to address important biological questions regarding the role of the A. gambiae mating plug. A second use for transglutaminase inhibitors exists for the study of haemolymph coagulation and immune responses to wound healing in insects.

【 授权许可】

Unknown   
© Le et al.; licensee BioMed Central Ltd. 2014. This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.