An Overview of The Phosphoproteomic Workflow

Öz

Proteomic studies permit the evaluation of pathological or physiological events in more detail through technological developments in the fields of mass spectrometry and chromatography. Protein phosphorylation, an important research area of mass spectrometry-based proteomic studies, plays an active role as a dynamic marker of molecular activity in the pathological alteration of cellular processes. Therefore, phosphoproteomic studies are gaining importance in the elucidation of molecular mechanisms of diseases and the development of effective diagnostic and treatment methods. Identification and quantitative analysis of phosphoproteins requires the use of highly efficient and reproducible methods. The development of existing methods is ongoing in order to maximize the efficiency of phosphoproteomic applications. We hope that this study, which explains the basic methodology of mass spectrometry-based phosphoproteomic research and discusses its practical challenges, will be useful for researchers interested in MS-based phosphoproteomics.

Anahtar Kelimeler

• Phosphoproteomics
• sample preparation
• enrichment
• quantitative phosphoproteomics
• mass spectrometry

Fosfoproteomik Uygulama Basamaklarına Genel Bakış

Öz

Proteomik çalışmalar, kütle spektrometresi ve kromatografi alanındaki teknolojik gelişmeler sayesinde patolojik ya da fizyolojik olayları daha detaylı olarak değerlendirebilmemize imkan sağlamaktadır. Kütle spektrometresi tabanlı proteomik çalışmaların önemli bir araştırma alanı olan protein fosforilasyonları, hücresel işleyişin patolojik değişiminde moleküler aktivitenin dinamik belirteçleri olarak etkin rol oynar. Bu nedenle hastalıkların moleküler mekanizmalarının aydınlatılmasında ve etkin tanı-tedavi yöntemlerinin geliştirilmesinde fosfoproteomik çalışmalar artan öneme sahiptir. Fosfoproteinlerin tanımlanması ve miktar analizlerinin yapılabilmesi oldukça verimli tekrarlanabilir yöntemlerin kullanılmasını gerektirir. Bu amaçla fosfoproteomik uygulamalardaki verimliliğin en üst düzeye çıkarılmasında mevcut yöntemler geliştirilmeye devam etmektedir. Kütle spektrometresi tabanlı fosfoproteomik araştırmaların temel metodolojisinin anlatıldığı ve uygulamalarda karşılaşılan zorlukların tartışıldığı bu çalışmanın, MS tabanlı fosfoproteomik alanına ilgi duyan araştırmacılara yardımcı olacağını umuyoruz.

Anahtar Kelimeler

• Fosfoproteomik
• örnek hazırlığı
• zenginleştirme
• niceliksel fosfoproteomik
• kütle spetrometresi

Kaynakça

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