Temperature scanning FTIR analysis of secondary structures of proteins embedded in amorphous sugar matrix

Imamura, Koreyoshi; Ohyama, Ken; Yokoyama, T.; Maruyama, Yoshinobu; Nakanishi, Koji

doi:10.1002/jps.21568

Cited by 18 publications

(11 citation statements)

References 32 publications

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“…It could be produced by Sphaerodes mycoparasitica interacting with Fusarium pathogen in MFR, as the trehalose seemed to enhance the efficacy of the biocontrol agent against pathogenic molds [48]. According to FTIR study conducted by Imamura et al [49], the trehalose embedded into protein matrix can also improve the cell membrane stability as measured by increased α-helix content (19%→45%), which was inversely correlated to β-sheet content (8%→4%), that aligned with results of this study.…”

Section: Estimation Of Protein Secondary Structures In Tripartite Systemsupporting

confidence: 88%

Specific Mycoparasite-Fusarium Graminearum Molecular Signatures in Germinating Seeds Disabled Fusarium Head Blight Pathogen’s Infection

Kim

Lahlali

Karunakaran

et al. 2021

IJMS

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Advances in Infrared (IR) spectroscopies have entered a new era of research with applications in phytobiome, plant microbiome and health. Fusarium graminearum 3-ADON is the most aggressive mycotoxigenic chemotype causing Fusarium head blight (FHB) in cereals; while Sphaerodes mycoparasitica is the specific Fusarium mycoparasite with biotrophic lifestyle discovered in cereal seeds and roots. Fourier transform infrared (FTIR) spectroscopy analyses depicted shifts in the spectral peaks related to mycoparasitism mainly within the region of proteins, lipids, also indicating a link between carbohydrates and protein regions, involving potential phenolic compounds. Especially, S. mycoparasitica contributes to significant changes in lipid region 3050–2800 cm−1, while in the protein region, an increasing trend was observed for the peaks 1655–1638 cm−1 (amide I) and 1549–1548 cm−1 (amide II) with changes in indicative protein secondary structures. Besides, the peak extending on the region 1520–1500 cm−1 insinuates a presence of aromatic compounds in presence of mycoparasite on the F. graminearum root sample. Monitoring shift in improved seed germination, fungus-fungus interface through scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM), and FTIR molecular signatures combined with principal component analysis (PCA) proved useful tools to detect an early mycoparasitism as a vital asset of the preventive biocontrol strategy against plant pathogens.

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Section: Estimation Of Protein Secondary Structures In Tripartite Systemsupporting

confidence: 88%

Specific Mycoparasite-Fusarium Graminearum Molecular Signatures in Germinating Seeds Disabled Fusarium Head Blight Pathogen’s Infection

Kim

Lahlali

Karunakaran

et al. 2021

IJMS

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“…The changes incurred by the secondary structural elements of BSA in aqueous solution were consistent with what other researchers have documented. 36,47 The modification in the secondary structure elements of BSA incorporated into Precirol AT05 using the wet granulation method is probably attributed to the interaction between dissolved BSA and Precirol AT05 hydrophobic surfaces and this increased as more lipid was introduced in the mixture. The addition of BSA as an aqueous solution to Precirol AT05 triggered its interactions with the lipid, which may not have occurred between solid BSA and lipid even by melting the lipid with BSA at an elevated temperature of 70 C for 1 h. This was perhaps due to the enhancement in protein adsorption and denaturation on lipid/hydrophobic surfaces upon addition in the solution state.…”

Section: Effects Of Incorporation Methods On Secondary Structure Of Bmentioning

confidence: 99%

Attenuated Total Reflection Fourier Transform Infrared (ATR FT-IR) Spectroscopy as an Analytical Method to Investigate the Secondary Structure of a Model Protein Embedded in Solid Lipid Matrices

et al. 2017

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Protein drugs may encounter conformational perturbations during the formulation processing of lipid-based solid dosage forms. In aqueous protein solutions, attenuated total reflection Fourier transform infrared (ATR FT-IR) spectroscopy can investigate these conformational changes following the subtraction of spectral interference of solvent with protein amide I bands. However, in solid dosage forms, the possible spectral contribution of lipid carriers to protein amide I band may be an obstacle to determine conformational alterations. The objective of this study was to develop an ATR FT-IR spectroscopic method for the analysis of protein secondary structure embedded in solid lipid matrices. Bovine serum albumin (BSA) was chosen as a model protein, while Precirol AT05 (glycerol palmitostearate, melting point 58 ℃) was employed as the model lipid matrix. Bovine serum albumin was incorporated into lipid using physical mixing, melting and mixing, or wet granulation mixing methods. Attenuated total reflection FT-IR spectroscopy and size exclusion chromatography (SEC) were performed for the analysis of BSA secondary structure and its dissolution in aqueous media, respectively. The results showed significant interference of Precirol ATO5 with BSA amide I band which was subtracted up to 90% w/w lipid content to analyze BSA secondary structure. In addition, ATR FT-IR spectroscopy also detected thermally denatured BSA solid alone and in the presence of lipid matrix indicating its suitability for the detection of denatured protein solids in lipid matrices. Despite being in the solid state, conformational changes occurred to BSA upon incorporation into solid lipid matrices. However, the extent of these conformational alterations was found to be dependent on the mixing method employed as indicated by area overlap calculations. For instance, the melting and mixing method imparted negligible effect on BSA secondary structure, whereas the wet granulation mixing method promoted more changes. Size exclusion chromatography analysis depicted the complete dissolution of BSA in the aqueous media employed in the wet granulation method. In conclusion, an ATR FT-IR spectroscopic method was successfully developed to investigate BSA secondary structure in solid lipid matrices following the subtraction of lipid spectral interference. The ATR FT-IR spectroscopy could further be applied to investigate the secondary structure perturbations of therapeutic proteins during their formulation development.

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“…Glass transition temperature (T g ) of the spin-dried drying solution (composition: see above) was determined using temperature ramp FTIR spectroscopy [15] . FTIR measurements were performed using a Nicolet Continuum FTIR Spectromicroscope (Thermo Electron Corporation LLC, Waltham, MA) equipped with an MCT micron detector.…”

Section: Methodsmentioning

confidence: 99%

Cryopreservation of Spin-Dried Mammalian Cells

et al. 2011

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This study reports an alternative approach to achieve vitrification where cells are pre-desiccated prior to cooling to cryogenic temperatures for storage. Chinese Hamster Ovary (CHO) cells suspended in a trehalose solution were rapidly and uniformly desiccated to a low moisture content (<0.12 g of water per g of dry weight) using a spin-drying technique. Trehalose was also introduced into the cells using a high-capacity trehalose transporter (TRET1). Fourier Transform Infrared Spectroscopy (FTIR) was used to examine the uniformity of water concentration distribution in the spin-dried samples. 62% of the cells were shown to survive spin-drying in the presence of trehalose following immediate rehydration. The spin-dried samples were stored in liquid nitrogen (LN2) at a vitrified state. It was shown that following re-warming to room temperature and re-hydration with a fully complemented cell culture medium, 51% of the spin-dried and vitrified cells survived and demonstrated normal growth characteristics. Spin-drying is a novel strategy that can be used to improve cryopreservation outcome by promoting rapid vitrification.

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Temperature scanning FTIR analysis of secondary structures of proteins embedded in amorphous sugar matrix

Cited by 18 publications

References 32 publications

Specific Mycoparasite-Fusarium Graminearum Molecular Signatures in Germinating Seeds Disabled Fusarium Head Blight Pathogen’s Infection

Specific Mycoparasite-Fusarium Graminearum Molecular Signatures in Germinating Seeds Disabled Fusarium Head Blight Pathogen’s Infection

Attenuated Total Reflection Fourier Transform Infrared (ATR FT-IR) Spectroscopy as an Analytical Method to Investigate the Secondary Structure of a Model Protein Embedded in Solid Lipid Matrices

Cryopreservation of Spin-Dried Mammalian Cells

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