Calcium-induced activity and folding of a repeat in toxin lipase from Antarctic Pseudomonas fluorescens strain AMS8

It is hypothesized that the Ca2+ 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...

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Main Authors: Mohd Ali, Nur Shidaa, Salleh, Abu Bakar, Raja Abdul Rahman, Raja Noor Zaliha, Leow, Thean Chor, Mohamad Ali, Mohd Shukuri
Format: Article
Language:English
Published: MDPI 2020
Online Access:http://psasir.upm.edu.my/id/eprint/38185/
http://psasir.upm.edu.my/id/eprint/38185/1/38185.pdf
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author Mohd Ali, Nur Shidaa
Salleh, Abu Bakar
Raja Abdul Rahman, Raja Noor Zaliha
Leow, Thean Chor
Mohamad Ali, Mohd Shukuri
author_facet Mohd Ali, Nur Shidaa
Salleh, Abu Bakar
Raja Abdul Rahman, Raja Noor Zaliha
Leow, Thean Chor
Mohamad Ali, Mohd Shukuri
author_sort Mohd Ali, Nur Shidaa
building UPM Institutional Repository
collection Online Access
description It is hypothesized that the Ca2+ 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 Ca2+ ion towards the activity and folding of AMS8 lipase through various biophysical characterizations. The results showed that CaCl2 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 CaCl2. Fluorescence spectroscopy analysis showed that the presence of CaCl2 increased protein folding and compactness. Dynamic light scattering (DLS) analysis suggested that AMS8 lipase became aggregated at a high concentration of CaCl2.The binding constant (Kd) value from the isothermal titration calorimetry (ITC) analysis proved that the Ca2+ ion was tightly bound to the AMS8 lipase. In conclusion, Ca2+ 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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spelling upm-381852020-05-03T23:05:04Z http://psasir.upm.edu.my/id/eprint/38185/ Calcium-induced activity and folding of a repeat in toxin lipase from Antarctic Pseudomonas fluorescens strain AMS8 Mohd Ali, Nur Shidaa Salleh, Abu Bakar Raja Abdul Rahman, Raja Noor Zaliha Leow, Thean Chor Mohamad Ali, Mohd Shukuri It is hypothesized that the Ca2+ 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 Ca2+ ion towards the activity and folding of AMS8 lipase through various biophysical characterizations. The results showed that CaCl2 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 CaCl2. Fluorescence spectroscopy analysis showed that the presence of CaCl2 increased protein folding and compactness. Dynamic light scattering (DLS) analysis suggested that AMS8 lipase became aggregated at a high concentration of CaCl2.The binding constant (Kd) value from the isothermal titration calorimetry (ITC) analysis proved that the Ca2+ ion was tightly bound to the AMS8 lipase. In conclusion, Ca2+ 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. MDPI 2020 Article PeerReviewed text en http://psasir.upm.edu.my/id/eprint/38185/1/38185.pdf Mohd Ali, Nur Shidaa and Salleh, Abu Bakar and Raja Abdul Rahman, Raja Noor Zaliha and Leow, Thean Chor and Mohamad Ali, Mohd Shukuri (2020) Calcium-induced activity and folding of a repeat in toxin lipase from Antarctic Pseudomonas fluorescens strain AMS8. Toxins, 12 (1). art. no. 27. pp. 1-14. ISSN 2072-6651 https://www.mdpi.com/2072-6651/12/1/27 10.3390/toxins12010027
spellingShingle Mohd Ali, Nur Shidaa
Salleh, Abu Bakar
Raja Abdul Rahman, Raja Noor Zaliha
Leow, Thean Chor
Mohamad Ali, Mohd Shukuri
Calcium-induced activity and folding of a repeat in toxin lipase from Antarctic Pseudomonas fluorescens strain AMS8
title Calcium-induced activity and folding of a repeat in toxin lipase from Antarctic Pseudomonas fluorescens strain AMS8
title_full Calcium-induced activity and folding of a repeat in toxin lipase from Antarctic Pseudomonas fluorescens strain AMS8
title_fullStr Calcium-induced activity and folding of a repeat in toxin lipase from Antarctic Pseudomonas fluorescens strain AMS8
title_full_unstemmed Calcium-induced activity and folding of a repeat in toxin lipase from Antarctic Pseudomonas fluorescens strain AMS8
title_short Calcium-induced activity and folding of a repeat in toxin lipase from Antarctic Pseudomonas fluorescens strain AMS8
title_sort calcium-induced activity and folding of a repeat in toxin lipase from antarctic pseudomonas fluorescens strain ams8
url http://psasir.upm.edu.my/id/eprint/38185/
http://psasir.upm.edu.my/id/eprint/38185/
http://psasir.upm.edu.my/id/eprint/38185/
http://psasir.upm.edu.my/id/eprint/38185/1/38185.pdf