Calcium-Induced Activity and Folding of a Repeat in Toxin Lipase from Antarctic Pseudomonas fluorescens Strain AMS8

It is hypothesized that the Ca 2+ ions were involved in the activity, folding and stabilization of many protein structures. Many of these proteins contain repeat in toxin (RTX) motifs. AMS8 lipase from Antarctic Pseudomonas fluorescens strain AMS8 was found to have three RTX motifs. So, this researc...

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Bibliographic Details
Published in:Toxins
Main Authors: Nur Shidaa Mohd Ali, Abu Bakar Salleh, Raja Noor Zaliha Raja Abd Rahman, Thean Chor Leow, Mohd Shukuri Mohamad Ali
Format: Article in Journal/Newspaper
Language:English
Published: MDPI AG 2020
Subjects:
rtx lipase
ams8 lipase
family i.3
rtx parallel β -roll motif repeat
ca 2+ ion
calcium binding
folding
activity
Medicine
R
Antarctic
Antarc*
Online Access:https://doi.org/10.3390/toxins12010027
https://doaj.org/article/e7c1a18d847c4fb8aa46448d5a99f4a9
Description
Summary:It is hypothesized that the Ca 2+ ions were involved in the activity, folding and stabilization of many protein structures. Many of these proteins contain repeat in toxin (RTX) motifs. AMS8 lipase from Antarctic Pseudomonas fluorescens strain AMS8 was found to have three RTX motifs. So, this research aimed to examine the influence of Ca 2+ ion towards the activity and folding of AMS8 lipase through various biophysical characterizations. The results showed that CaCl 2 increased lipase activity. The far-UV circular dichroism (CD) and Fourier-transform infrared (FTIR) analysis suggested that the secondary structure content was improved with the addition of CaCl 2 . Fluorescence spectroscopy analysis showed that the presence of CaCl 2 increased protein folding and compactness. Dynamic light scattering (DLS) analysis suggested that AMS8 lipase became aggregated at a high concentration of CaCl 2 .The binding constant (K d ) value from the isothermal titration calorimetry (ITC) analysis proved that the Ca 2+ ion was tightly bound to the AMS8 lipase. In conclusion, Ca 2+ ions play crucial roles in the activity and folding of the AMS8 lipase. Calcium binding to RTX nonapeptide repeats sequences will induced the formation and folding of the RTX parallel β -roll motif repeat structure.

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