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The impact of ultrafiltration on the molecular properties of bovine lactoferrin: molecular mass, primary structure and glycosylation variations

Version 1.0

Dyrda-Terniuk, Tetiana; Pomastowski, Paweł, 2024, "The impact of ultrafiltration on the molecular properties of bovine lactoferrin: molecular mass, primary structure and glycosylation variations", https://doi.org/10.18150/TPOVHF, RepOD, V1

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Description

Mass spectra of intact bovine lactoferrin before and after ultrafiltration obtained by matrix-assisted laser desorption/ionization time-of-flight mass (MALDI-TOF-MS) spectrometer Bruker ultrafleXtreme (Bruker Daltonik, Bremen, Germany) were recorded in the positive ion linear mode in m/z range of 25000-250000 using accelerating voltage of 25 kV. Saturated solution of sinapinic acid (SA) dissolved in TA30 (mixture of 0.1 % TFA and ACN at the 70:30 (v/v) was used as matrix solution, while Protein Calibration Standard II (Bruker Daltonics, Bremen) was selected for calibration.

Peptide mass fingerprint (PMF) of bovine lactoferrin before and after ultrafiltration in both, reduced and non-reduced conditions, was recorded in MS mode in the reflectron positive ion mode within m/z range of 500-3500. Saturated α-cyano-4-hydroxycinnamic acid (HCCA) in TA30 was used as a matrix solution, while the Peptide Calibration Standard II (Bruker Daltonics, Bremen) was applied as a calibrant.

MS/MS fragments of bovine lactoferrin were obtained in Laser Ionization Fragmentation Technologies (LIFT) mode. The common criteria selected for the bLF identification was the peak intensity ˃ 1000 a.u., except for bovine lactoferrin before ultrafiltration at non-reduced conditions in which case the criteria was changed to intensity ˃ 3000 a.u. Cysteine carbamidomethylation and methionine oxidation were selected as global and variable modifications, respectively. Variable modification, such as HexNAc (N), Hex(1)HexNAc(2) (N), HexNAc(2)dHex(1) (N), HexNAc(2)dHex(1) (N), Hex(1)HexNAc(1)dHex(1) (N), and Hex(1)HexNAc(1)NeuAc(1) (N) were applied in order to indicate matches (glycopeptides), representing the potential glycosylation sites. Mascot database along with BioTools were applied for the result interpretation.

(2024-07-03)
Subject
Chemistry
Keyword

bovine lactoferrin, ultrafiltration, identification, MALDI, peptide mass fingerprint, LIFT

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Intact bovine lactoferrin_spectra.zip
ZIP Archive - 520.9 KB - Jul 3, 2024 - 3 Downloads
MD5: fbaec455690e02cb3dfc02ed0d9bfab3
License: CC BY - Creative Commons Attribution 4.0
Folder contains MALDI-TOF-MS spectra of intact bovine lactoferrin before and after ultrafiltration.
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Peptide mass fingerprint_bovine lactoferrin.zip
ZIP Archive - 4.1 MB - Jul 3, 2024 - 0 Downloads
MD5: 79fbda0ab676aaf763c8585597f71b6e
License: CC BY - Creative Commons Attribution 4.0
Folder contains data set regarding protein identification in MS mode: matched, unmatched peptides, digest score, sequence, intensity coverage and PMF specta of bovine lactoferrin before and after ultrafiltration performed in both, reduced and non-reduced conditions.
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Identification data after fragmentation in LIFT mode_bovine lactoferrin.zip
ZIP Archive - 11.2 MB - Jul 3, 2024 - 0 Downloads
MD5: d7c1328cabf5162bdf397b3d89a81c16
License: CC BY - Creative Commons Attribution 4.0
Folder contains data set regarding bovine lactoferrin identification in LIFT MS/MS mode (The common criteria selected for the bLF identification was the peak intensity ˃ 1000 a.u., except for bovine lactoferrin before ultrafiltration at non-reduced conditions in which case the criteria was changed to intensity ˃ 3000 a.u.) and list of matched MS/MS fragment ions.
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Glycosylation sites in bovine lactoferrin.zip
ZIP Archive - 83.3 KB - Jul 3, 2024 - 2 Downloads
MD5: eb35d42b924f07df930b159b98be8527
License: CC BY - Creative Commons Attribution 4.0
Folder contains data set regarding identification of potential glycosites of bovine lactoferrin before and after ultrafiltration performed in both, reduced and non-reduced conditions by using the series of variable modifications, such as HexNAc (N), Hex(1)HexNAc(2) (N), HexNAc(2)dHex(1) (N), HexNAc(2)dHex(1) (N), Hex(1)HexNAc(1)dHex(1) (N), and Hex(1)HexNAc(1)NeuAc(1) (N).
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readme.txt
Plain Text - 5.5 KB - Jul 3, 2024 - 1 Download
MD5: 74a7c5811cfd0252dbed2ca145dc4d42
License: CC0 Creative Commons Zero 1.0
This readme file was generated on 2024-07-02 by Tetiana Dyrda-Terniuk
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