کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
2020991 1069222 2010 5 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Expression of four mutant fibrinogen γC domains in Pichia pastoris confirms them as causes of hypofibrinogenaemia
موضوعات مرتبط
علوم زیستی و بیوفناوری بیوشیمی، ژنتیک و زیست شناسی مولکولی زیست شیمی
پیش نمایش صفحه اول مقاله
Expression of four mutant fibrinogen γC domains in Pichia pastoris confirms them as causes of hypofibrinogenaemia
چکیده انگلیسی
Mutations in the fibrinogen gene cluster can cause low plasma fibrinogen concentrations, known as hypofibrinogenaemia. It is important to verify whether a detected sequence variant in this cluster is deleterious or benign and this can be accomplished using protein expression systems. In this study, four mutations in the fibrinogen γC domain that had previously been described in patients with hypofibrinogenaemia were introduced into a γC construct and expressed in a Pichia pastoris yeast system to investigate their effects on protein stability and secretion. These experiments showed that the fibrinogen Middlemore (N230D), Dorfen (A289V), Mannheim II (H307Y), and Muncie (T371I) mutations were not secreted, supporting their causative role in hypofibrinogenaemia. Overexpression of the N230D, A289V and H307Y mutants revealed that the majority of the synthesised protein was retained in the endoplasmic reticulum, with only a minor proportion reaching the trans-Golgi network. Regardless, none of this protein was secreted which confirms that the four mutations investigated are indeed responsible for hypofibrinogenaemia.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Protein Expression and Purification - Volume 73, Issue 2, October 2010, Pages 184-188
نویسندگان
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