Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2017 Oct 27;7:14234. doi: 10.1038/s41598-017-14074-9

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

© The Author(s) 2017

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

PMC Copyright notice

Analysis of Cab1 pantothenate kinase activity. (A–C) Cab1 enzyme activity as a function of ATP consumption (A and B) or ADP production (C). The amount of ATP in the reaction was measured using the Kinase-Glo Plus Luminescent Kinase (Kinase-Glo) assay. In this reaction, the luminescence signal (Relative light units- RLU) is inversely correlated with kinase activity. Kinase-Glo Reagent was added following a kinase reaction and incubated for 60 min at room temperature. Reactions were carried out in the presence or absence of ATP, PA and Cab1, and luminescence determined using a BioTek SynergyMx Microplate reader (A). Heat inactivated Cab1 (Denat Cab1) is used as a control (B). Cab1 activity as a function of ADP production was measured using the ADP-Glo Kinase assay. In this reaction, luminescence (RLU) is directly correlated with the kinase activity. Kinase reactions were carried out in the presence of active or heat inactivated Cab1 (C). Data are expressed as the mean ± SE of four experiments. (D) Mass spectrometry analysis of Cab1 pantothenate phosphoprylation activity. Reactions were conducted in the presence of PA and ATP and in the presence (+Cab1) or absence (-Cab1) of Cab1. Upper panels show negative ion scans specifically looking for the mass of PA-1 (mass range 217–219). The lower panels show negative ion scans specifically looking for the mass of P-PA-1 (mass range 297–299). The y-axis was normalized to 5000 units for comparison purposes.