MALDI-TOF mass spectrometric identification of novel intercellular space peptides

Ильина, А. П.; Куликова, О. Г.; Maltsev, Dmitry I; Krasnov, M. S.; Rybakova, E. Yu.; Skripnikova, V. S.; Кузнецова, Е. С.; Buryak, А. К.; Yamskova, V. P.; Yamskov, I. A.

doi:10.1134/s0003683811020049

Cited by 6 publications

(5 citation statements)

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“…Las secuencias de los péptidos inicialmente diseñados y de sus análogos se presentan en la Tabla 1. Las modificaciones en la secuencia de los péptidos GIBIM-P2 a GIBIM-P5, usadas para diseñar los análogos peptídicos, buscaban aumentar la carga neta positiva, la naturaleza hidrófoba y el porcentaje de probabilidad de ser un péptido antimicrobiano, propiedades fisicoquímicas directamente relacionadas con la actividad antimicrobiana (Il'ina, et al, 2011;Bakshi, et al, 2014). Por esta razón, el diseño de los péptidos análogos GIBIM-P3A6K, GIBIM-P5T2K y GIBIM-P5S9K, y los residuos de alanina (A), treonina (T) y serina (S) se reemplazaron por lisina (K) para aumentar el carácter catiónico de los péptidos (Teixeira, et al, 2012;Sengupta, et al, 2008;Vila-Farrés, et al, 2012).…”

Section: Resultados Y Discusiónunclassified

“…En cuanto a la purificación del péptido antimicrobiano sintetizado, el grado de pureza se determina usualmente mediante cromatografía liquida de alta resolución en fase reversa (reversed phase high-performance liquid chromatography, RP-HPLC) (Aguilar, 2004). Para la determinación de la masa molecular del péptido, se utilizan la espectrometría de masas (mass spectometry, MS), dada su sensibilidad, velocidad y alto grado de especificidad molecular (Il'ina, et al, 2011), y el dicroísmo circular (DC), técnica alternativa de espectroscopía de absorción electrónica, que permite determinar la estructura secundaria del péptido. Hoy existen métodos de deconvolución que utilizan bases de datos y permiten establecer una estructura muy acertada (Bakshi, et al, 2014).…”

Section: Introductionunclassified

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Diseño, síntesis, caracterización y evaluación in vitro de la actividad de los péptidos antimicrobianos contra bacterias patógenas resistentes a antibióticos

López

2019

Rev. Acad. Colomb. Cienc. Ex. Fis. Nat.

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Los péptidos antimicrobianos han atraído mucha atención como nuevos agentes terapéuticos contra enfermedades infecciosas. En este estudio se hizo el diseño racional in silico de 18 péptidos catiónicos con actividad antimicrobiana contra bacterias patógenas resistentes utilizando el programa DEPRAMP desarrollado en el Grupo de Investigación en Bioquímica y Microbiología de la Universidad Industrial de Santander. Posteriormente, los péptidos diseñados se sintetizaron en fase sólida con el método de 9-fluorenilmetoxicarbonilo en medio ácido. Se obtuvieron secuencias cortas de 17 aminoácidos con un grado de pureza entre 95 y 98 %, estructura secundaria de hélice alfa, carga neta catiónica (entre +3 y +6), punto isoeléctrico entre 10,04 y 12,03 e índice de hidropatía entre -0,62 y 1,14. Todos los péptidos antimicrobianos mostraron actividad antibacteriana y bactericida in vitro frente al menos una de las cepas patógenas estudiadas: Escherichia coli O157: H7, Pseudomonas aeruginosa y Staphylococcus aureus resistente a la meticilina. Los péptidos antimicrobianos GIBIM-P5S9K y GIBIM-P5F8W registraron la mejor actividad antibacteriana, alcanzando una concentración mínima inhibitoria (CMI 99) en rangos de 0,5 a 25 μM frente a las tres cepas evaluadas, de las cuales Escherichia coli O157: H7 fue la más sensible frente al péptido antimicrobiano GIBIMP5F8W, con una CMI 99 de 0,5 μM y una concentración mínima bactericida de 10 μM, en tanto que la cepa de Pseudomonas aeruginosa fue la más resistente, con una CMI de más de 100 μM frente a más de cinco péptidos antimicrobianos. La toxicidad de los péptidos sobre los eritrocitos produjo un porcentaje de hemólisis menor al 40 % en concentraciones de 50 μM. Por su parte, en las líneas celulares de carcinoma de pulmón A549 y HepG2, el único compuesto que presentó toxicidad fue GIBIM-P5F8W, presentando un 36% de células viables en concentraciones de 100 μM del péptido en la línea celular A549.

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Section: Resultados Y Discusiónunclassified

Section: Introductionunclassified

Diseño, síntesis, caracterización y evaluación in vitro de la actividad de los péptidos antimicrobianos contra bacterias patógenas resistentes a antibióticos

López

2019

Rev. Acad. Colomb. Cienc. Ex. Fis. Nat.

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“…However, there is no signal with m/z 4404 detected in our prior LC-MS peptidomics work on tissue extracts and releasates. [41] Il’ina et al [43] identified a group of peptides in Wistar rat brain extract, referred to as membrane-tropic homeostatic tissue-specific bioregulators, including a peptide at m/z 4403; unfortunately, the primary sequence of the peptide was not reported, and so this remains unknown. High-mass resolution MALDI MS measurements of the extracts were performed using a Bruker SolariX FT-ICR mass spectrometer.…”

Section: Resultsmentioning

confidence: 99%

Optically Guided Single Cell Mass Spectrometry of Rat Dorsal Root Ganglia to Profile Lipids, Peptides and Proteins

et al. 2018

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The mammalian dorsal root ganglia (DRG) are located on the dorsal roots of the spinal nerves and contain cell bodies of primary sensory neurons. DRG cells have been classified into subpopulations based on their size, morphology, intracellular markers, response to stimuli, and neuropeptides. To understand the connections between DRG chemical heterogeneity and cellular function, we performed optically guided, high-throughput single cell profiling using sequential matrix-assisted laser desorption/ionization mass spectrometry (MS) to detect lipids, peptides, and several proteins in individual DRG cells. Statistical analysis of the resulting mass spectra allows stratification of the DRG population according to cellular morphology and, presumably, major cell types. A subpopulation of small cells contained myelin proteins, which are abundant in Schwann cells, and mass spectra of several larger cells contained peaks matching neurofilament, vimentin, myelin basic protein S, and thymosin beta proteins. Of the over 1000 cells analyzed, approximately 78 % produced putative peptide-rich spectra, allowing the population to be classified into three distinct cell types. Two signals with m/z 4404 and 5487 were exclusively observed in a cell type, but could not be matched to results of our previous liquid chromatography-MS analyses.

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“…The speed of movement and, accordingly, the time it takes to travel from the point of ionization to the detector, is inversely proportional to the mass of the ions. Knowing the path length of ion movement from the ionizer to the detector, as well as the time of this movement, it is possible to calculate the speed of ion movement and, based on its value, calculate the mass of particles present in the analyte, as well as generate a spectrum characterizing the qualitative composition of the object under study [25].…”

Section: Maldi-tofmethodsmentioning

confidence: 99%

Improvement of Enzymatic Saccharification of Cellulose-Containing Raw Materials Using Aspergillus niger

et al. 2021

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Enzymatic hydrolysis of cellulose-containing raw materials, using Aspergillus niger, were studied. Filter paper, secondary cellulose-containing or starch-containing raw materials, miscanthus cellulose after alkaline or acid pretreatment, and wood chip cellulose, were used as substrates. The study focused on a wild A. niger strain, treated, or not (control), by ultraviolet (UV) irradiations for 45, 60, or 120 min (UV45, UV60, or UV120), or by UV irradiation for 120 min followed by a chemical treatment with NaN3 + ItBr for 30 min or 80 min (UV120 + CH30 or UV120 + CH80). A mixture of all the A. niger strains (MIX) was also tested. A citrate buffer, at 50 mM, wasthe most suitable for enzymatic hydrolysis. As the UV exposure time increased to 2 h, the cellulase activity of the surviving culturewas increased (r = 0.706; p < 0.05). The enzymatic activities of the obtained strains, towards miscanthus cellulose, wood chips, and filter paper, were inferior to those obtained with commercial enzymes (8.6 versus 9.1 IU), in some cases. Under stationary hydrolysis at 37 °C, pH = 4.7, the enzymatic activity of A. niger UV120 + CH30 was 24.9 IU. The enzymatic hydrolysis of secondary raw materials, using treated A. niger strains, was themost effective at 37 °C. Similarly, the most effective treatment of miscanthus cellulose and wood chips occurred at 50 °C. The maximum conversion of cellulose to glucose was observed using miscanthus cellulose (with alkaline pretreatment), and the minimum conversion was observed when using wood chips. The greatest value of cellulase activity was evidenced in the starch-containing raw materials, indicating that A. niger can ferment not only through cellulase activity, but also via an amylolytic one.

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MALDI-TOF mass spectrometric identification of novel intercellular space peptides

Cited by 6 publications

References 1 publication

Diseño, síntesis, caracterización y evaluación in vitro de la actividad de los péptidos antimicrobianos contra bacterias patógenas resistentes a antibióticos

Diseño, síntesis, caracterización y evaluación in vitro de la actividad de los péptidos antimicrobianos contra bacterias patógenas resistentes a antibióticos

Optically Guided Single Cell Mass Spectrometry of Rat Dorsal Root Ganglia to Profile Lipids, Peptides and Proteins

Improvement of Enzymatic Saccharification of Cellulose-Containing Raw Materials Using Aspergillus niger

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