Local conformations affect the histidine tag-Ni<sup>2+</sup> binding affinity of BinA and BinB proteins

Sudarat Tharad; Chontida Tangsongcharoen; Panadda Boonserm; José L. Toca-Herrera; Kanokporn Srisucharitpanit; Sudarat Tharad; Chontida Tangsongcharoen; Panadda Boonserm; José L. Toca-Herrera; Kanokporn Srisucharitpanit

doi:10.3934/biophy.2020011

AIMS Biophysics

2020, Volume 7, Issue 3: 133-143. doi: 10.3934/biophy.2020011

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Local conformations affect the histidine tag-Ni²⁺ binding affinity of BinA and BinB proteins

1.
Institute for Biophysics, Department of Nanobiotechnology, University of Natural Resources and Life Sciences (BOKU), Vienna 1190, Austria
2.
Faculty of Allied Health Sciences, Burapha University, Chonburi, Thailand 20131
3.
Institute of Molecular Biosciences, Mahidol University, Nakhon Pathom, Thailand 73170

Received: 20 March 2020 Accepted: 12 May 2020 Published: 19 May 2020

Binary toxin (Bin) is one of the bio-larvicidal toxin produced by Lysinibacillus sphaericus. Two component proteins, BinA and BinB toxins are required simultaneously to exert its larvicidal activity. The binary toxin has been proposed to act initially on the susceptible cell membrane. Here, the cell membrane binding of the binary toxin was imitated via specific histidine (His)-nickel ion (Ni²⁺) chelating. The N-terminal His-conjugated binary toxins (His-Bin) were attached onto the Ni²⁺-lipid bilayer surface besides its facilitating of purification process. The N-terminal conjugation of histidine tag did not interfere with the folding structure of both toxins. Subsequently, the attachment of binary toxins on the lipid membrane was successful with Ni²⁺-phosphatidylcholine (POPC)/phosphatidylethanolamine (POPE) bilayers (a model membrane that mimics the mosquito cell membrane) but not for Ni²⁺-phosphatidylcholine bilayer. However, His-BinA formed unstable attachment with Ni²⁺-POPC/POPE bilayers since it could be removed by buffer rinsing. In contrast, His-BinB required imidazole solution to detach from Ni²⁺-POPC/POPE. Particularly, His-BinB had higher binding affinity to Ni²⁺-ion than His-BinA. The lipid membrane attachment led to the initial finding that although BinA and BinB toxins share high homology structures, their capability for Ni²⁺ chelation was different. The local N-terminal structure of binary toxin seems to interfere the His-Ni²⁺ chelating of His-BinA.
- Lysinibacillus sphaericus,
- binary toxin,
- histidine tagged protein,
- nickel ion chelating,
- quartz crystal microbalance with dissipation (QCM-D)
Citation: Sudarat Tharad, Chontida Tangsongcharoen, Panadda Boonserm, José L. Toca-Herrera, Kanokporn Srisucharitpanit. Local conformations affect the histidine tag-Ni2+ binding affinity of BinA and BinB proteins[J]. AIMS Biophysics, 2020, 7(3): 133-143. doi: 10.3934/biophy.2020011

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Abstract

Binary toxin (Bin) is one of the bio-larvicidal toxin produced by Lysinibacillus sphaericus. Two component proteins, BinA and BinB toxins are required simultaneously to exert its larvicidal activity. The binary toxin has been proposed to act initially on the susceptible cell membrane. Here, the cell membrane binding of the binary toxin was imitated via specific histidine (His)-nickel ion (Ni²⁺) chelating. The N-terminal His-conjugated binary toxins (His-Bin) were attached onto the Ni²⁺-lipid bilayer surface besides its facilitating of purification process. The N-terminal conjugation of histidine tag did not interfere with the folding structure of both toxins. Subsequently, the attachment of binary toxins on the lipid membrane was successful with Ni²⁺-phosphatidylcholine (POPC)/phosphatidylethanolamine (POPE) bilayers (a model membrane that mimics the mosquito cell membrane) but not for Ni²⁺-phosphatidylcholine bilayer. However, His-BinA formed unstable attachment with Ni²⁺-POPC/POPE bilayers since it could be removed by buffer rinsing. In contrast, His-BinB required imidazole solution to detach from Ni²⁺-POPC/POPE. Particularly, His-BinB had higher binding affinity to Ni²⁺-ion than His-BinA. The lipid membrane attachment led to the initial finding that although BinA and BinB toxins share high homology structures, their capability for Ni²⁺ chelation was different. The local N-terminal structure of binary toxin seems to interfere the His-Ni²⁺ chelating of His-BinA.

Acknowledgments

The authors would like to kindly thanks Monruedee Srisaisap (Mahidol University) and Jacqueline Friedmann (BOKU) for technical supporting. This work was supported by the Thailand Research Fund (TRF, MRG5980076), the Austrian Science Fund (FWF) (grant number P29562-N28), and the faculty of Allied Health Sciences, Burapha University.

Conflict of interest

The authors declare no conflicts of interest.

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