Site specific PEGylation of β-lactoglobulin at glutamine residues and its influence on conformation and antigenicity

Luo, Shuhong; Lu, Xiao-San; Liu, Chengmei; Zhong, Junzhen; Zhou, Lei; Chen, Tingting

doi:10.1016/j.foodres.2019.05.038

Cited by 11 publications

(6 citation statements)

References 51 publications

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“…When conjugating a PEG‐maleimide to Cys232, the disulfide bond between Cys232 to the additional cysteine cap no longer exists, thus its quenching should not be present anymore, causing the increase of the intrinsic fluorescence intensity. Other studies show that PEGylation can decrease the protein intrinsic fluorescence 38 or has no effect 39–41 . The decrease of intrinsic fluorescence in β‐lactoglobulin (β‐LG) after PEGylation was explained as a combined action of the shielding effect of the PEG chain and the exposure of the Trp residues to the solvent upon PEGylation 38 .…”

Section: Discussionmentioning

confidence: 99%

“…The decrease of intrinsic fluorescence in β‐lactoglobulin (β‐LG) after PEGylation was explained as a combined action of the shielding effect of the PEG chain and the exposure of the Trp residues to the solvent upon PEGylation 38 . In the case of granulocyte colony‐stimulating factor (G‐CSF), PEGylation at two different sites (Lys41 and Gln135) resulted in unchanged or increased intrinsic fluorescence intensity, respectively 38 …”

Section: Discussionmentioning

confidence: 99%

“…[39][40][41] The decrease of intrinsic fluorescence in β-lactoglobulin (β-LG) after PEGylation was explained as a combined action of the shielding effect of the PEG chain and the exposure of the Trp residues to the solvent upon PEGylation. 38 In the case of granulocyte colonystimulating factor (G-CSF), PEGylation at two different sites (Lys41 and Gln135) resulted in unchanged or increased intrinsic fluorescence intensity, respectively. 38 PEGylated AAT displays the same biological activity as AAT, indicating that PEGylation does not affect the structure of the most important part of this protein, the reaction center loop (RCL).…”

Section: Effects Of Pegylation On the Secondary And Tertiary Structur...mentioning

confidence: 99%

“…38 In the case of granulocyte colonystimulating factor (G-CSF), PEGylation at two different sites (Lys41 and Gln135) resulted in unchanged or increased intrinsic fluorescence intensity, respectively. 38 PEGylated AAT displays the same biological activity as AAT, indicating that PEGylation does not affect the structure of the most important part of this protein, the reaction center loop (RCL). 32 These results are in line with Zhu et al 42 and Cantin et al 33 It is reported from several studies that PEGylation does not alter the tertiary structure of a protein substantially.…”

Section: Effects Of Pegylation On the Secondary And Tertiary Structur...mentioning

confidence: 99%

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Impact of the PEG length and PEGylation site on the structural, thermodynamic, thermal, and proteolytic stability of mono‐PEGylated alpha‐1 antitrypsin

et al. 2022

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Conjugation to polyethylene glycol (PEG) is a widely used approach to improve the therapeutic value of proteins essentially by prolonging their body residence time. PEGylation may however induce changes in the structure and/or the stability of proteins and thus on their function(s). The effects of PEGylation on the thermodynamic stability can either be positive (stabilization), negative (destabilization), or neutral (no effect). Moreover, various factors such as the PEG length and PEGylation site can influence the consequences of PEGylation on the structure and stability of proteins. In this study, the effects of PEGylation on the structure, stability, and polymerization of alpha1‐antitrypsin (AAT) were investigated, using PEGs with different lengths, different structures (linear or 2‐armed) and different linking chemistries (via amine or thiol) at two distinct positions of the sequence. The results show that whatever the size, position, and structure of PEG chains, PEGylation (a) does not induce significant changes in AAT structure (either at the secondary or tertiary level); (b) does not alter the stability of the native protein upon both chemical‐ and heat‐induced denaturation; and (c) does not prevent AAT to fully refold and recover its activity following chemical denaturation. However, the propensity of AAT to aggregate upon heat treatment was significantly decreased by PEGylation, although PEGylation did not prevent the irreversible inactivation of the enzyme. Moreover, conjugation to PEG, especially 2‐armed 40 kDa PEG, greatly improved the proteolytic resistance of AAT. PEGylation of AAT could be a promising strategy to prolong its half‐life after infusion in AAT‐deficient patients and thereby decrease the frequency of infusions.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Effects Of Pegylation On the Secondary And Tertiary Structur...mentioning

confidence: 99%

Section: Effects Of Pegylation On the Secondary And Tertiary Structur...mentioning

confidence: 99%

See 2 more Smart Citations

Impact of the PEG length and PEGylation site on the structural, thermodynamic, thermal, and proteolytic stability of mono‐PEGylated alpha‐1 antitrypsin

et al. 2022

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“…Prior to the measurement, samples were diluted to 1 mg/mL of protein concentration with ultra-pure water. The emission spectrum of tryptophan was recorded from 300 to 400 nm with a slit width of 3 nm, after excitation at 280 nm [18].…”

Section: And Tryptophan Fluorescence Spectroscopymentioning

confidence: 99%

Characteristics and Functional Properties of Maillard Reaction Products from α-Lactalbumin and Polydextrose

Dai

Wang

Luo

et al. 2023

Foods

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The characteristics and the functions of Maillard reaction products (MRPs) produced by polydextrose (PD), a new type of prebiotic, and α-lactalbumin (α-LA) were valued. PD and α-LA were incubated at 60 °C and 79% relative humidity for up to 72 h to prepare MRPs. The results showed that the absorbance and fluorescence intensity of heated α-LA-PD increased, and the amount of free amino groups reduced as the reaction progressed, which confirmed the formation of different stages of MRPs. Electrophoresis revealed an increase in molecular mass and the degree of covalent cross-linking. The secondary structure of MRPs experienced no significant changes with the measurement of circular dichroism (CD), while the tertiary structure gradually unfolded, exposing hydrophobic groups. Furthermore, a significant increase was detected in the radical-scavenging activity of 2,2-diphenyl-1-picrylhydrazyl (DPPH) and the ferric reducing/antioxidant power (FRAP) of MRPs. The findings offer a foundation for understanding the structural and functional features of MRPs in formula milk powder.

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A new site-specific monoPEGylated β-lactoglobulin at the N-terminal: Effect of different molecular weights of mPEG on its conformation and antigenicity

Luo

Zhou

et al. 2021

Food Chemistry

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Site specific PEGylation of β-lactoglobulin at glutamine residues and its influence on conformation and antigenicity

Cited by 11 publications

References 51 publications

Impact of the PEG length and PEGylation site on the structural, thermodynamic, thermal, and proteolytic stability of mono‐PEGylated alpha‐1 antitrypsin

Impact of the PEG length and PEGylation site on the structural, thermodynamic, thermal, and proteolytic stability of mono‐PEGylated alpha‐1 antitrypsin

Characteristics and Functional Properties of Maillard Reaction Products from α-Lactalbumin and Polydextrose

A new site-specific monoPEGylated β-lactoglobulin at the N-terminal: Effect of different molecular weights of mPEG on its conformation and antigenicity

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