Microcalorimetric Studies of the Interactions of Lysozyme with Immobilized Metal Ions: Effects of Ion, pH Value, and Salt Concentration

Lin, Fu‐Yang; Chen, Wen-Yih; Sang, Lung-Ching

doi:10.1006/jcis.1999.6193

Cited by 44 publications

(23 citation statements)

References 35 publications

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“…In addition, several literature (6,17,(43)(44)(45)(46) have demonstrated that the adsorption affinities and retention of proteins fused with histidine tags toward immobilized metal ions increased with the number of exposed histidines. Since the heat required for the dehydration processes (i.e., (a) and (b) processes) did not mostly attribute to the difference in values of H ads as compared to the coordination formation process (i.e., the (c) process) under the conditions studied (21), and the accompanied enthalpy change of hydration process caused by the binding-induced structural change of the protein may have been only a minor contribution to the total binding enthalpy, if any, the major contribution to the difference in H ads values between SjGST and SjGST/His with Ni-NTA gels, therefore, was concluded to be the heat revealed by the process of the formation of coordination compounds (i.e., the (c) process).…”

Section: Effects Of the Hexa-histidine Tagmentioning

confidence: 94%

“…Moreover, because the pH value (pH 8) taken in this study was higher than the pK a value of the imidazole group (pK a ∼ 6.5), the formation of coordination bonds dominated the adsorption process (21) (21), suggesting that the heat revealed at the formation of coordination bonds between SjGST/His and either metal ion (i.e., the (c) process) was not significantly different. The theory of hard and soft acids and bases (HSAB) purposed by also suggests that the coordination of the nitrogen atom with Ni 2+ is as thermodynamically stable as that with Co 2+ because both of them are intermediate type of ions.…”

Section: Thermodynamic Characterization Of the Interaction Mechanismsmentioning

confidence: 94%

“…However, for highly exposed histidyl residues adjacent on the affinity tag of SjGST/His under a denaturing condition, the difference in binding topography between two IMAC adsorbents with SjGST/His was considered to be minor. Therefore, based on the same denaturing condition and the postulation (similar binding topography), the main difference of H ads for Ni-NTA and TALON with SjGST/His was possibly resulting from the difference in the immobilized metal ions, leading to different heats required for dehydration (i.e., (a) and (b) processes) and revealed by the formation of coordination bonds (i.e., the (c) process) (21).…”

Section: Thermodynamic Characterization Of the Interaction Mechanismsmentioning

confidence: 98%

“…The because the dehydration process of the adsorbents and the coordination interaction and/or nonspecific interaction formed between SjGST/His and the immobilized metal ions (i.e., (b) and (c) processes) are affected by the immobilized metal ions (21). Moreover, because the pH value (pH 8) taken in this study was higher than the pK a value of the imidazole group (pK a ∼ 6.5), the formation of coordination bonds dominated the adsorption process (21) (21), suggesting that the heat revealed at the formation of coordination bonds between SjGST/His and either metal ion (i.e., the (c) process) was not significantly different.…”

Section: Thermodynamic Characterization Of the Interaction Mechanismsmentioning

confidence: 99%

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Microcalorimetric Study of the Effect of Hexa-histidine Tag and Denaturant on the Interaction Mechanism between Protein and Metal-Chelating Gel

Lin

Chen

2001

Journal of Colloid and Interface Science

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Section: Effects Of the Hexa-histidine Tagmentioning

confidence: 94%

Section: Thermodynamic Characterization Of the Interaction Mechanismsmentioning

confidence: 94%

Section: Thermodynamic Characterization Of the Interaction Mechanismsmentioning

confidence: 98%

Section: Thermodynamic Characterization Of the Interaction Mechanismsmentioning

confidence: 99%

See 2 more Smart Citations

Microcalorimetric Study of the Effect of Hexa-histidine Tag and Denaturant on the Interaction Mechanism between Protein and Metal-Chelating Gel

Lin

Chen

2001

Journal of Colloid and Interface Science

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“…The fourth theory was proposed by Lin et al (Lin et al 2000), based on the calorimetric studies on the interactions of imidazole and lysozyme (Chen et al 1997, Lin et al 1999 In this chapter, HIC was explored to remove the 70-kDa contaminant from GSTpurified modular VP1 capsomeres (Section 6. (Yamniuk et al 2013).…”

Section: Removal Of Gst Tag and Undigested Gst-tagged Modular Vp1 Usimentioning

confidence: 99%

Engineering of virus-like particles for alternative vaccine candidate targeting a hypervariable peptide antigen element

Anggraeni¹

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A promising alternative to replace the current egg-or cell culture-based technology for vaccine production from live viruses is virus-like particle (VLP) technology based on a microbial platform. VLPs are macromolecular assemblies of viral capsid proteins, which have been shown to tolerate insertion of antigen modules via genetic recombinant technology, yielding modular VLPs. Many studies on modular VLPs presume that when a peptide antigen element is taken out from the intact proteins and then modularised on VLPs, it is unable to fold into its native structure. However, until now, presentation of a peptide antigen element on a VLP and the impact of the display strategy to present the antigen element on the quality of the resulting antibodies (i.e. the ability of the antibodies to recognise the intact protein) are not fully understood. This thesis aims to understand the underlying fundamentals regarding modularisation of peptide antigen elements on VLPs for induction of high-quality antibodies. A hypervariable receptor-binding domain, Helix 190 (H190), from the hemagglutinin protein of influenza A virus was used as a model for modularisation on VLPs from murine polyomavirus (MuPyV) VP1 protein. Four major findings are presented. Firstly, two display strategies, i.e. arraying of H190 in tandem repeats and the use of helix promoter elements, were shown to display H190 in its immunogenic form equally. However, modularisation using tandem repeat display induced antibodies of a higher quality than modularisation using helix promoter elements.Secondly, the quality of antibodies induced by the tandem repeat display bearing two copies of H190 was optimum, thus no significant improvement was observed following the use of adjuvant or increasing the copy number of H190. Additionally, the increase in the copy number of H190 was shown to reduce the assembly capability and solubility of modular VP1 in an environment that was optimised for wild-type VP1. Thirdly, this thesis shows the novel finding in the use of flanking ionic elements to stabilise VLP precursors, termed as capsomeres, bearing two copies of H190 containing a hydrophobic stretch, which caused aggregation. Fourthly, the first steps towards obtaining the atomic crystal structure of presented H190 on a modular protein were performed, i.e. a mild and satisfactory laboratory process was developed to achieve high-purity modular VP1 capsomeres, unattainable using previously established expression and purification process of wild-type MuPyV VP1. This thesis shows a step forward towards understanding the presentation of a peptide antigen element on a VLP that enables induction of highquality antibodies, and towards VLP engineering to manipulate the aggregation and solubility of modular VP1. VLP technology based on a microbial platform presented here is iii a potentially safe and effective alternative vaccine candidate that targets a hypervariable peptide antigen element. The speed of the microbial platform allows a rapid response to the hypervariability of the pe...

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Recent trends and some applications of isothermal titration calorimetry in biotechnology

Roselin

Lin

et al. 2010

Biotechnology Journal

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Isothermal titration calorimeters (ITCs) are thermodynamic instruments used for the determination of enthalpy changes in any physical/chemical reaction. This can be applied in various fields of biotechnology. This review explains ITC applications, especially in bioseparation, drug development and cell metabolism. In liquid chromatography, the separation/purification of specific proteins or polypeptides in a mixture is usually achieved by varying the adsorption affinities of the different proteins/polypeptides for the adsorbent under different mobile-phase conditions and temperatures. Using ITC analysis, the binding mechanism of proteins with adsorbent solid material is derived by elucidating enthalpy and entropy changes, which offer valuable guidelines for designing experimental conditions in chromatographic separation. The binding affinity of a drug with its target is studied by deriving binding enthalpy and binding entropy. To improve the binding affinity, suitable lead compounds for a drug can be identified and their affinity tested by ITC. Recently ITC has also been used in studying cell metabolism. The heat produced by animal cells in culture can be used as a primary indicator of the kinetics of cell metabolism, which provides key information for drug bioactivity and operation parameters for process cell culture.

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Microcalorimetric Studies of the Interactions of Lysozyme with Immobilized Metal Ions: Effects of Ion, pH Value, and Salt Concentration

Cited by 44 publications

References 35 publications

Microcalorimetric Study of the Effect of Hexa-histidine Tag and Denaturant on the Interaction Mechanism between Protein and Metal-Chelating Gel

Microcalorimetric Study of the Effect of Hexa-histidine Tag and Denaturant on the Interaction Mechanism between Protein and Metal-Chelating Gel

Engineering of virus-like particles for alternative vaccine candidate targeting a hypervariable peptide antigen element

Recent trends and some applications of isothermal titration calorimetry in biotechnology

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