1. Thymosin fraction 5 re-evaluated after 35 years by high-resolution mass spectrometry.
- Author
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Hannappel E, Iavarone F, and Castagnola M
- Subjects
- Animals, Chromatography, High Pressure Liquid, Drug Stability, Drug Storage, Freeze Drying, Humans, Protein Precursors analysis, Protein Precursors isolation & purification, Proteomics methods, Thymosin analysis, Thymosin chemistry, Thymosin isolation & purification, Time Factors, Ubiquitin analysis, Ubiquitin isolation & purification, Mass Spectrometry methods, Thymosin analogs & derivatives
- Abstract
Objectives: We reevaluated a lyophilized sample of thymosin fraction 5, stored for 37 years at room temperature, by high-resolution mass spectrometry in terms of stability and yet uncharacterized polypeptides that could be biological important substances., Methods: A top-down proteomic platform based on high-performance liquid chromatography (HPLC) coupled to high-resolution LTQ-Orbitrap mass spectrometry (MS) was applied to molecular characterization of polypeptides present in thymosin fraction 5., Results: We detected more than 100 monoisotopic masses corresponding to thymosin β4 and truncated forms of ubiquitin, prothymosin α, thymosin β4, and thymosin β9. Additionally, we discovered a new polypeptide present in thymosin fraction 5 and identified it as intact SH3 domain-binding glutamic acid-rich-like protein 3., Conclusion: In spite of the well-known proteolytic processes inherent to the preparation of thymosin fraction 5, still uncharacterized polypeptides as well as truncated forms of already well-known thymosins are present in fraction 5 after long-term storage. Therefore, continuing characterization of thymosin fraction 5 is even nowadays highly promising.
- Published
- 2018
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