کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
1192288 1492307 2011 8 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Top-down high-resolution electron capture dissociation mass spectrometry for comprehensive characterization of post-translational modifications in Rhesus monkey cardiac troponin I
موضوعات مرتبط
مهندسی و علوم پایه شیمی شیمی آنالیزی یا شیمی تجزیه
پیش نمایش صفحه اول مقاله
Top-down high-resolution electron capture dissociation mass spectrometry for comprehensive characterization of post-translational modifications in Rhesus monkey cardiac troponin I
چکیده انگلیسی

Top-down high-resolution electron capture dissociation (ECD) mass spectrometry (MS) is a powerful approach for comprehensive characterization of large proteins with labile post-translational modifications (PTMs). Phosphorylation of cardiac troponin I (cTnI) is a prominent determinant of cardiac function. Non-human primates (NHPs) are valuable models for biomedical research due to their genetic similarity to humans. To address the emerging need for in-depth understanding of NHP heart models, we have applied top-down high-resolution tandem mass spectrometry in conjugation with immunoaffinity chromatography purification to comprehensively characterize NHP cTnI. Our data revealed that cTnI affinity purified from NHP Rhesus monkey hearts was N-terminally acetylated and mono- or bis-phosphorylated. ECD unambiguously identified Ser22/Ser23 as the only basally phosphorylated sites with a phosphorylation order between these two sites (Ser23 phosphorylates prior to Ser22) which is different from those previously reported for mouse and human cTnI. Our study also strongly supports the nonergodic mechanism of ECD since no neutral loss of phosphates was observed in ECD spectra. Taken together, top-down MS with ECD is extremely valuable for studying labile PTMs in large proteins, which adds critical knowledge to our understanding of protein PTM regulations in health and disease.

.Figure optionsDownload high-quality image (171 K)Download as PowerPoint slideResearch highlights▶ Top-down ECD MS was employed for comprehensively charactering PTMs in monkey cTnI. ▶ Rhesus monkey cTnI was N-terminally acetylated and mono- or bis-phosphorylated. ▶ Top-down ECD MS unambiguously identified Ser22/Ser23 as the only basally phosphorylated sites. ▶ The phosphorylation order was determined: Ser23 phosphorylates prior to Ser22. ▶ No neutral phosphate loss in ECD spectra supports the nonergodic mechanism of ECD.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: International Journal of Mass Spectrometry - Volume 305, Issues 2–3, 15 August 2011, Pages 95–102
نویسندگان
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