Xử lý proteolytic của đơn vị alpha11.2 kênh Ca2+ loại L trong tế bào thần kinh

F1000Research - Tập 6 - Trang 1166
Olivia R. Buonarati1, Peter B. Henderson1, Geoffrey G. Murphy2, Mary C. Horne1, Johannes Hell1
1Department of Pharmacology, University of California, Davis, CA
2Department of Molecular and Integrative Physiology, Molecular and Behavioral Neuroscience Institute, University of Michigan, Ann Arbor, MI

Tóm tắt

Bối cảnh: Kênh Ca2+ loại L Cav1.2 là một yếu tố điều chỉnh nổi bật trong tính kích thích của tế bào thần kinh, tính dẻo của synap và biểu hiện gen. Yếu tố trung tâm của Cav1.2 là đơn vị α11.2 tạo lỗ. Đơn vị này tồn tại dưới hai dạng kích thước chính, mà khối lượng phân tử của chúng đã được chứng minh là khó xác định chính xác. Công trình nghiên cứu gần đây gợi ý rằng α11.2 bị cắt đứt proteolytic ở vị trí giữa miền tạo lỗ thứ hai và thứ ba trong tổng số bốn miền tạo lỗ của nó (Michailidiset al, 2014).Phương pháp: Để xác định khối lượng phân tử rõ ràng (MR) của các dạng kích thước α11.2, một loạt các thí nghiệm Western blot hệ thống trên mô não và cả dạng đầy đủ lẫn dạng α11.2 bị cắt ngắn ở phần C-terminus được tiến hành sử dụng sáu kháng thể đặc hiệu vùng khác nhau chống lại α11.2.Kết quả: Dạng đầy đủ của α11.2 di chuyển như mong đợi, với MR rõ ràng ~250 kDa. Một dạng ngắn hơn có độ phổ biến tương đương với MR rõ ràng ~210 kDa chỉ có thể được phát hiện trong các blots miễn dịch được kiểm tra bằng các kháng thể nhận biết α11.2 tại một epitope 400 hoặc nhiều hơn residues phía thượng lưu của C-terminus.Kết luận: Hai dạng kích thước chính của α11.2 là dạng đầy đủ và một dạng ngắn hơn, thiếu ~350 residues tách biệt ở C-terminus. Việc cắt đứt giữa kênh như được gợi ý bởi Michailidiset al (2014) là rất ít xảy ra trong mô não.

Từ khóa


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