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Đặc trưng hóa sự tương tác giữa Protein LIM liên kết với Actinin (ALP) và miền que của α-actinin
Tóm tắt
Protein PDZ-LIM là một gia đình các adaptor tín hiệu tương tác với protein liên kết chéo actin, α-actinin, thông qua các miền PDZ của chúng hoặc qua các vùng nội bộ giữa các miền PDZ và LIM. Ba trong số các protein PDZ-LIM có một động thái ZM 26-residue được bảo tồn trong vùng nội bộ, nhưng cấu trúc của vùng nội bộ thì chưa được biết đến. Trong nghiên cứu này, sử dụng phương pháp quang phổ phân cực tròn và cộng hưởng từ hạt nhân (NMR), chúng tôi đã chỉ ra rằng vùng nội bộ ALP (các residue 107–273) chủ yếu là không có cấu trúc rõ ràng trong dung dịch, nhưng có khả năng tương tác với miền que α-actinin trong ống nghiệm, và đồng định vị với α-actinin trên các sợi căng thẳng trong cơ thể sống. Phân tích NMR cho thấy rằng việc titrat ALP với miền que α-actinin dẫn đến sự ổn định của ALP. Một peptide tổng hợp (các residue 175–196) chứa nửa đầu N- của động thái ZM đã được phát hiện là tương tác trực tiếp với miền que α-actinin trong các phép đo phản ứng bề mặt plasmon (SPR). Những vị trí thiếu hụt ngắn ở hoặc trước động thái ZM đã làm mất đi sự định vị của ALP trên các sợi căng thẳng actin. Vùng nội bộ của ALP có vẻ chủ yếu là không có cấu trúc nhưng vẫn có chức năng. Động thái ZM định nghĩa một phần bề mặt tương tác giữa ALP và miền que α-actinin.
Từ khóa
Tài liệu tham khảo
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