Cargando…

Reprogramming fatty acyl specificity of lipid kinases via C1 domain engineering

C1 domains are lipid-binding modules that regulate membrane activation of kinases, nucleotide exchange factors, and other C1-containing proteins to trigger signal transduction. Despite annotation of typical C1 domains as diacylglycerol (DAG) and phorbol ester sensors, the function of atypical counte...

Descripción completa

Detalles Bibliográficos
Autores principales: Ware, Timothy B., Franks, Caroline E., Granade, Mitchell E., Zhang, Mingxing, Kim, Kee-Beom, Park, Kwon-Sik, Gahlmann, Andreas, Harris, Thurl E., Hsu, Ku-Lung
Formato: Online Artículo Texto
Lenguaje:English
Publicado: 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7117826/
https://www.ncbi.nlm.nih.gov/pubmed/31932721
http://dx.doi.org/10.1038/s41589-019-0445-9
_version_ 1783514450000609280
author Ware, Timothy B.
Franks, Caroline E.
Granade, Mitchell E.
Zhang, Mingxing
Kim, Kee-Beom
Park, Kwon-Sik
Gahlmann, Andreas
Harris, Thurl E.
Hsu, Ku-Lung
author_facet Ware, Timothy B.
Franks, Caroline E.
Granade, Mitchell E.
Zhang, Mingxing
Kim, Kee-Beom
Park, Kwon-Sik
Gahlmann, Andreas
Harris, Thurl E.
Hsu, Ku-Lung
author_sort Ware, Timothy B.
collection PubMed
description C1 domains are lipid-binding modules that regulate membrane activation of kinases, nucleotide exchange factors, and other C1-containing proteins to trigger signal transduction. Despite annotation of typical C1 domains as diacylglycerol (DAG) and phorbol ester sensors, the function of atypical counterparts remains ill-defined. Here, we assign a key role for atypical C1 domains in mediating DAG fatty acyl specificity of diacylglycerol kinases (DGKs) in live cells. Activity-based proteomics mapped C1 probe binding as a principal differentiator of type 1 DGK active sites that combined with global metabolomics revealed a role for C1s in lipid substrate recognition. Protein engineering by C1 domain swapping demonstrated that exchange of typical and atypical C1s is functionally tolerated and can directly program DAG fatty acyl specificity of type 1 DGKs. Collectively, we describe a protein engineering strategy for studying metabolic specificity of lipid kinases to assign a role for atypical C1 domains in cell metabolism.
format Online
Article
Text
id pubmed-7117826
institution National Center for Biotechnology Information
language English
publishDate 2020
record_format MEDLINE/PubMed
spelling pubmed-71178262020-07-13 Reprogramming fatty acyl specificity of lipid kinases via C1 domain engineering Ware, Timothy B. Franks, Caroline E. Granade, Mitchell E. Zhang, Mingxing Kim, Kee-Beom Park, Kwon-Sik Gahlmann, Andreas Harris, Thurl E. Hsu, Ku-Lung Nat Chem Biol Article C1 domains are lipid-binding modules that regulate membrane activation of kinases, nucleotide exchange factors, and other C1-containing proteins to trigger signal transduction. Despite annotation of typical C1 domains as diacylglycerol (DAG) and phorbol ester sensors, the function of atypical counterparts remains ill-defined. Here, we assign a key role for atypical C1 domains in mediating DAG fatty acyl specificity of diacylglycerol kinases (DGKs) in live cells. Activity-based proteomics mapped C1 probe binding as a principal differentiator of type 1 DGK active sites that combined with global metabolomics revealed a role for C1s in lipid substrate recognition. Protein engineering by C1 domain swapping demonstrated that exchange of typical and atypical C1s is functionally tolerated and can directly program DAG fatty acyl specificity of type 1 DGKs. Collectively, we describe a protein engineering strategy for studying metabolic specificity of lipid kinases to assign a role for atypical C1 domains in cell metabolism. 2020-01-13 2020-02 /pmc/articles/PMC7117826/ /pubmed/31932721 http://dx.doi.org/10.1038/s41589-019-0445-9 Text en Users may view, print, copy, and download text and data-mine the content in such documents, for the purposes of academic research, subject always to the full Conditions of use:http://www.nature.com/authors/editorial_policies/license.html#terms
spellingShingle Article
Ware, Timothy B.
Franks, Caroline E.
Granade, Mitchell E.
Zhang, Mingxing
Kim, Kee-Beom
Park, Kwon-Sik
Gahlmann, Andreas
Harris, Thurl E.
Hsu, Ku-Lung
Reprogramming fatty acyl specificity of lipid kinases via C1 domain engineering
title Reprogramming fatty acyl specificity of lipid kinases via C1 domain engineering
title_full Reprogramming fatty acyl specificity of lipid kinases via C1 domain engineering
title_fullStr Reprogramming fatty acyl specificity of lipid kinases via C1 domain engineering
title_full_unstemmed Reprogramming fatty acyl specificity of lipid kinases via C1 domain engineering
title_short Reprogramming fatty acyl specificity of lipid kinases via C1 domain engineering
title_sort reprogramming fatty acyl specificity of lipid kinases via c1 domain engineering
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7117826/
https://www.ncbi.nlm.nih.gov/pubmed/31932721
http://dx.doi.org/10.1038/s41589-019-0445-9
work_keys_str_mv AT waretimothyb reprogrammingfattyacylspecificityoflipidkinasesviac1domainengineering
AT frankscarolinee reprogrammingfattyacylspecificityoflipidkinasesviac1domainengineering
AT granademitchelle reprogrammingfattyacylspecificityoflipidkinasesviac1domainengineering
AT zhangmingxing reprogrammingfattyacylspecificityoflipidkinasesviac1domainengineering
AT kimkeebeom reprogrammingfattyacylspecificityoflipidkinasesviac1domainengineering
AT parkkwonsik reprogrammingfattyacylspecificityoflipidkinasesviac1domainengineering
AT gahlmannandreas reprogrammingfattyacylspecificityoflipidkinasesviac1domainengineering
AT harristhurle reprogrammingfattyacylspecificityoflipidkinasesviac1domainengineering
AT hsukulung reprogrammingfattyacylspecificityoflipidkinasesviac1domainengineering