Endpoint kinetic assay at variable pH using mouse Acidic Mammalian Chitinase

This directory contains all files required to analyze the mouse AMCase enzyme kinetics across pH 2.0 to 8.0 presented in the manuscript Díaz et al., bioRxiv (2023).

Enzyme kinetics were performed using a Tecan Spark multimode microplate reader and data was analyzed using Graphpad Prism. Figures were compiled using Adobe Illustrator.

Files included in this directory:

20210906 - mAMCase_CatD_6xHis (pmRED006)

- 20210907 - Purification contains FPLC chromatograms and corresponding SDS-PAGE gel stained with InstantBlue.

- 202110XX _pHX_100nM contains four replicates performed at that specific pH using 100 nM mAMCase.

- 384w_exp.toml contains a 384-well plate layout of the experimental conditions. This file is compatible with wellmap.

- 384w_std.toml contains a 384-well plate layout of the 4MU standards. This file is compatible with wellmap.

Figures
- contains PDFs of V_max, k_cat, K_M, catalytic efficiency, and the initial rate for all pH points tested. 

20210906 - mAMCase_CatD_6xHis (pmRED006) - Data.pzfx

- contains all data from 20210906 - mAMCase_CatD_6xHis (pmRED006), including standard curves, initial time points, and initial rates curves. 

20210906 - mAMCase_CatD_6xHis (pmRED006) - Summary.pzfx

- contains summary data of kinetic parameters V_max, k_cat, K_M, catalytic efficiency, and initial rates for all pH points tested.

Contact:
Roberto Efraín Díaz, robertoefrain.diaz@ucsf.edu

James Fraser, jfraser@fraserlab.com