High-resolution mass spectrometry for thymosins detection and characterization

Cabras, Tiziana; Iavarone, Federica; Martelli, Claudia; Delfino, Daniela; Rossetti, Diana Valeria; Inserra, Ilaria; Manconi, Barbara; Desiderio, Claudia; Messana, Irene; Hannappel, Ewald; Faa, Gavino; Castagnola, Massimo

doi:10.1517/14712598.2015.1009887

Cited by 8 publications

(4 citation statements)

References 43 publications

(59 reference statements)

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“…14 Our observation is consistent with a previous work, which also noted that a TDP method resulted in higher sequence coverage compared to a BUP method for T β 4 and T β 10 proteins. 19…”

Section: Resultsmentioning

confidence: 99%

“…14 Our observation is consistent with a previous work, which also noted that a TDP method resulted in higher sequence coverage compared to a BUP method for Tβ4 and Tβ10 proteins. 19 For J9P127 (Tβ4) and its proteoforms a total of seven peptides were detected including proteolytic fragments (Table S2). The predominant proteoform was formed by removal of the N-terminal Met (a 131 Da mass loss) and N-terminal acetylation of the second amino acid Ser (a 42 Da mass gain) resulting in overall a decrease in protein mass by 89 Da (Table 1 and Figure 2).…”

Section: Characterization Of Proteoforms and Endogenous Peptides By E...mentioning

confidence: 99%

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Top-down mass spectrometry for characterizing the low molecular weight proteome of canine osteosarcoma cell phenotypes

Yang,

Stanisheuski,

Song

et al. 2023

Eur J Mass Spectrom (Chichester)

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Osteosarcoma (OS) is the most common primary malignant tumor of bone, which occupying about 20% of all bone cancers. To increase understanding of the biology of OS, we developed and evaluated a top-down mass spectrometry approach to detect, identify and quantify low molecular weight (MW) proteins (i.e., 1 kDa < MW < 30 kDa) in osteosarcoma cells. Top-down proteomic (TDP) data was acquired using reversed phase nano-liquid chromatography in conjunction with high-resolution mass spectrometry and resulted in the assignment of 328 proteins and 820 proteoforms or degradation products with high confidence. Eight post-translational modifications (PTMs) were identified in the present study, including N-terminal acetylation, lysine acetylation, succinylation, malonylation, serine/tyrosine phosphorylation, histidine methylation and N-acetylleucine. We confirmed that a truncated N-terminal proteoform lost 73 Da of mass through removal of the N-terminal Met (−131 Da), acetylation of the second amino acid (+42 Da), and Met oxidation (+16 Da). The results showed that the levels of proteoforms/biodegradable peptides correlated with the metastatic phenotypes of osteosarcoma cell lines. This study demonstrates the benefits of TDP for the characterization and relative quantification of proteoforms with relevance to OS biology and the potential of small molecular weight proteoforms to serve as a still underappreciated source of biomarkers.

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“…14 Our observation is consistent with a previous work, which also noted that a TDP method resulted in higher sequence coverage compared to a BUP method for T β 4 and T β 10 proteins. 19…”

Section: Resultsmentioning

confidence: 99%

Section: Characterization Of Proteoforms and Endogenous Peptides By E...mentioning

confidence: 99%

Top-down mass spectrometry for characterizing the low molecular weight proteome of canine osteosarcoma cell phenotypes

Yang,

Stanisheuski,

Song

et al. 2023

Eur J Mass Spectrom (Chichester)

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“…Conversely, the inhibition of osteogenic differentiation towards promotion of the adipogenic one has also been reported in mesenchymal stem cells . The Tβ4 e Tβ10 C‐terminal truncated form have been already characterized by our group in different tissues; however, their biological role is still unclear .…”

Section: Resultsmentioning

confidence: 99%

Lipoaspirate fluid proteome: A preliminary investigation by LC-MS top-down/bottom-up integrated platform of a high potential biofluid in regenerative medicine

et al. 2016

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The lipoaspirate fluid (LAF) is emerging as a potentially valuable source in regenerative medicine. In particular, our group recently demonstrated that it is able to exert osteoinductive properties in vitro. This original observation stimulated the investigation of the proteomic component of LAF, by means of LC-ESI-LTQ-Orbitrap-MS top-down/bottom-up integrated approach, which represents the object of the present study. Top-down analyses required the optimization of sample pretreatment procedures to enable the correct investigation of the intact proteome. Bottom-up analyses have been directly applied to untreated samples after monodimensional SDS-PAGE separation. The analysis of the acid-soluble fraction of LAF by top-down approach allowed demonstrating the presence of albumin and hemoglobin fragments (i.e. VV- and LVV-hemorphin-7), thymosins β4 and β10 peptides, ubiquitin and acyl-CoA binding protein; adipogenesis regulatory factor, perilipin-1 fragments, and S100A6, along with their PTMs. Part of the bottom-up proteomic profile was reproducibly found in both tested samples. The bottom-up approach allowed demonstrating the presence of proteins, listed among the components of adipose tissue and/or comprised within the ASCs intracellular content and secreted proteome. Our data provide a first glance on the LAF molecular profile, which is consistent with its tissue environment. LAF appeared to contain bioactive proteins, peptides and paracrine factors, suggesting its potential translational exploitation.

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“…The analysis of the undigested proteome provided a new perspective and better understanding of the naturally occurring fragmentome of AC tissue: of particular interest was the description of hemoglobin-derived peptides, C-terminal truncated forms of ubiquitin, and beta-thymosins peptides, all previously characterized in pediatric tumors of the posterior cranial fossa [23, 42] and of unclear biological significance in these diseases.…”

Section: Discussionmentioning

confidence: 99%

Investigating the Protein Signature of Adamantinomatous Craniopharyngioma Pediatric Brain Tumor Tissue: Towards the Comprehension of Its Aggressive Behavior

et al. 2019

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Although histologically benign, adamantinomatous craniopharyngioma (AC) pediatric brain tumor is a locally aggressive disease that frequently determines symptoms and hormonal dysfunctions related to the mass effect on the surrounding structures. Another typical feature of this benign neoplasm is the presence of voluminous liquid cysts frequently associated with the solid component. Even if studies have been devoted to the proteomic characterization of the tumor intracystic fluid, poor explorations have been performed on its solid part, principally investigated by transcriptomics technologies. In the present study, seven specimens of AC whole tumor tissue have been analyzed by LC-MS for a preliminary assessment of the proteomic profile by a top-down/bottom-up integrated approach. Thymosin beta 4, ubiquitin, calmodulin, S100 proteins, prothymosin α isoform 2, alpha-defensins 1-4, and fragments largely belonging to vimentin, hemoglobin, and glial fibrillary acidic protein characterized the intact proteome. The identification of alpha-defensins, formerly characterized in AC intracystic fluid, reinforces the hypothesis of a role for inflammation in tumor pathogenesis. A total number of 1798 unique elements were identified by a bottom-up approach with a special focus on the 433 proteins commonly characterized in the 85.7% of the samples analyzed. Their gene ontology classification evidenced the involvement of the adherence system, intermediate filaments, and actin cytoskeleton in tumor pathogenesis and of elements part of the Wnt, FGF, and EGFR signaling pathways. In addition, proteins involved in calcium modulation, innate immunity, inflammation, CCKR and integrin signaling, and gonadotropin-releasing hormone receptor pathways were also outlined. Further than confirming proteomic data previously obtained on AC intracystic fluid, these results offer a preliminary overview of the AC whole tissue protein phenotype, adding new hints towards the comprehension of this still obscure pediatric brain tumor.

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High-resolution mass spectrometry for thymosins detection and characterization

Abstract: The clinical and prognostic significance and the origin of these proteoforms have to be deeply investigated.

Cited by 8 publications

References 43 publications

Top-down mass spectrometry for characterizing the low molecular weight proteome of canine osteosarcoma cell phenotypes

Top-down mass spectrometry for characterizing the low molecular weight proteome of canine osteosarcoma cell phenotypes

Lipoaspirate fluid proteome: A preliminary investigation by LC-MS top-down/bottom-up integrated platform of a high potential biofluid in regenerative medicine

Investigating the Protein Signature of Adamantinomatous Craniopharyngioma Pediatric Brain Tumor Tissue: Towards the Comprehension of Its Aggressive Behavior

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