دورية أكاديمية
MS Identification of Blood Plasma Proteins Concentrated on a Photocrosslinker-Modified Surface.
| العنوان: | MS Identification of Blood Plasma Proteins Concentrated on a Photocrosslinker-Modified Surface. |
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| المؤلفون: | Gordeeva AI; Institute of Biomedical Chemistry (IBMC), 119121 Moscow, Russia., Valueva AA; Institute of Biomedical Chemistry (IBMC), 119121 Moscow, Russia., Rybakova EE; Institute of Biomedical Chemistry (IBMC), 119121 Moscow, Russia., Ershova MO; Institute of Biomedical Chemistry (IBMC), 119121 Moscow, Russia., Shumov ID; Institute of Biomedical Chemistry (IBMC), 119121 Moscow, Russia., Kozlov AF; Institute of Biomedical Chemistry (IBMC), 119121 Moscow, Russia., Ziborov VS; Institute of Biomedical Chemistry (IBMC), 119121 Moscow, Russia., Kozlova AS; Institute of Biomedical Chemistry (IBMC), 119121 Moscow, Russia., Zgoda VG; Institute of Biomedical Chemistry (IBMC), 119121 Moscow, Russia., Ivanov YD; Institute of Biomedical Chemistry (IBMC), 119121 Moscow, Russia., Ilgisonis EV; Institute of Biomedical Chemistry (IBMC), 119121 Moscow, Russia., Kiseleva OI; Institute of Biomedical Chemistry (IBMC), 119121 Moscow, Russia., Ponomarenko EA; Institute of Biomedical Chemistry (IBMC), 119121 Moscow, Russia., Lisitsa AV; Institute of Biomedical Chemistry (IBMC), 119121 Moscow, Russia., Archakov AI; Institute of Biomedical Chemistry (IBMC), 119121 Moscow, Russia., Pleshakova TO; Institute of Biomedical Chemistry (IBMC), 119121 Moscow, Russia. |
| المصدر: | International journal of molecular sciences [Int J Mol Sci] 2023 Dec 28; Vol. 25 (1). Date of Electronic Publication: 2023 Dec 28. |
| نوع المنشور: | Journal Article |
| اللغة: | English |
| بيانات الدورية: | Publisher: MDPI Country of Publication: Switzerland NLM ID: 101092791 Publication Model: Electronic Cited Medium: Internet ISSN: 1422-0067 (Electronic) Linking ISSN: 14220067 NLM ISO Abbreviation: Int J Mol Sci Subsets: MEDLINE |
| أسماء مطبوعة: | Original Publication: Basel, Switzerland : MDPI, [2000- |
| مواضيع طبية MeSH: | Proteomics* , Blood Proteins*, Humans ; Aluminum Silicates ; Mass Spectrometry |
| مستخلص: | This work demonstrates the use of a modified mica to concentrate proteins, which is required for proteomic profiling of blood plasma by mass spectrometry (MS). The surface of mica substrates, which are routinely used in atomic force microscopy (AFM), was modified with a photocrosslinker to allow "irreversible" binding of proteins via covalent bond formation. This modified substrate was called the AFM chip. This study aimed to determine the role of the surface and crosslinker in the efficient concentration of various types of proteins in plasma over a wide concentration range. The substrate surface was modified with a 4-benzoylbenzoic acid N-succinimidyl ester (SuccBB) photocrosslinker, activated by UV irradiation. AFM chips were incubated with plasma samples from a healthy volunteer at various dilution ratios (10 2 X, 10 4 X, and 10 6 X). Control experiments were performed without UV irradiation to evaluate the contribution of physical protein adsorption to the concentration efficiency. AFM imaging confirmed the presence of protein layers on the chip surface after incubation with the samples. MS analysis of different samples indicated that the proteomic profile of the AFM-visualized layers contained common and unique proteins. In the working series of experiments, 228 proteins were identified on the chip surface for all samples, and 21 proteins were not identified in the control series. In the control series, a total of 220 proteins were identified on the chip surface, seven of which were not found in the working series. In plasma samples at various dilution ratios, a total of 146 proteins were identified without the concentration step, while 17 proteins were not detected in the series using AFM chips. The introduction of a concentration step using AFM chips allowed us to identify more proteins than in plasma samples without this step. We found that AFM chips with a modified surface facilitate the efficient concentration of proteins owing to the adsorption factor and the formation of covalent bonds between the proteins and the chip surface. The results of our study can be applied in the development of highly sensitive analytical systems for determining the complete composition of the plasma proteome. |
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| معلومات مُعتمدة: | 075-15-2021-933 The Ministry of Education and Science of the Russian Federation |
| فهرسة مساهمة: | Keywords: atomic force microscopy; crosslinker; mass spectrometry; protein immobilization |
| المشرفين على المادة: | V8A1AW0880 (mica) 0 (Blood Proteins) 0 (Aluminum Silicates) |
| تواريخ الأحداث: | Date Created: 20240111 Date Completed: 20240112 Latest Revision: 20240113 |
| رمز التحديث: | 20240113 |
| مُعرف محوري في PubMed: | PMC10778900 |
| DOI: | 10.3390/ijms25010409 |
| PMID: | 38203578 |
| قاعدة البيانات: | MEDLINE |
Full Text Finder | تدمد: | 1422-0067 |
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| DOI: | 10.3390/ijms25010409 |