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Gán đồng bộ và chính xác tự động các tần số protein cho NMR trạng thái rắn
Tóm tắt
Quá trình gán đồng bộ đại diện cho một nút thắt thời gian tiêu tốn và có nguy cơ sai sót trong các nghiên cứu cấu trúc của protein bằng NMR trạng thái rắn (ssNMR). Phần mềm tự động hóa quá trình này do đó đang được quan tâm cao. Các quy trình được phát triển trong vài thập kỷ qua cho NMR ở trạng thái dung dịch không thể áp dụng trực tiếp cho ssNMR do chất lượng dữ liệu vốn dĩ thấp hơn gây ra bởi độ nhạy thấp hơn và các đường phổ rộng hơn, dẫn đến chồng chéo giữa các đỉnh. Gần đây, những nỗ lực đầu tiên hướng tới các quy trình đặc biệt dành cho ssNMR đã được thực hiện (Schmidt et al. trong J Biomol NMR 56(3):243–254, 2013). Tại đây, chúng tôi trình bày một phương pháp tự động mạnh mẽ, có thể gán chính xác các tần số protein bằng cách sử dụng danh sách đỉnh từ một tập hợp nhỏ các thí nghiệm ssNMR 2D và 3D đơn giản, có thể áp dụng trong các trường hợp có độ nhạy thấp. Phương pháp này được chứng minh trên ba sinh chất sinh học được đánh dấu đồng nhất với 13C, 15N với những thách thức khác nhau trong việc gán. Cụ thể, đối với miền gán kết immunoglobulin B1 của protein G do liên cầu khuẩn, việc gán tự động cho thấy độ chính xác 100% cho các tần số xương sống và 91,8% khi bao gồm tất cả các carbon chuỗi bên. Nó được chứng minh, bằng cách sử dụng quy trình tạo ra quang phổ nhân tạo với chiều rộng đường tăng dần, rằng phương pháp của chúng tôi, GAMES_ASSIGN có thể xử lý một lượng lớn các đỉnh chồng chéo trong việc gán. Tác động của việc bao gồm các thí nghiệm ssNMR khác nhau cũng được đánh giá.
Từ khóa
#NMR trạng thái rắn #gán đồng bộ protein #tự động hóa #độ chính xác #quang phổ nhân tạoTài liệu tham khảo
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