1FS3

CRYSTAL STRUCTURE OF WILD-TYPE BOVINE PANCREATIC RIBONUCLEASE A

1FS3 の概要

• エントリーDOI: 10.2210/pdb1fs3/pdb
• 関連するPDBエントリー: 1DP1 1EIC 1EID 1EIE
• 分子名称: Ribonuclease A (2 entities in total)
• 機能のキーワード: ribonuclease, rnase a, bovine pancreas, hydrolase
• 由来する生物種: Bos taurus (cattle)
• 細胞内の位置: Secreted: P61823
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 13708.33

構造登録者

Chatani, E.,Hayashi, R.,Moriyama, H.,Ueki, T. (登録日: 2000-09-08, 公開日: 2002-02-13, 最終更新日: 2024-10-30)

主引用文献

Chatani, E.,Hayashi, R.,Moriyama, H.,Ueki, T.
Conformational strictness required for maximum activity and stability of bovine pancreatic ribonuclease A as revealed by crystallographic study of three Phe120 mutants at 1.4 A resolution.
Protein Sci., 11:72-81, 2002

PubMed Abstract: The replacement of Phe120 with other hydrophobic residues causes a decrease in the activity and thermal stability in ribonuclease A (RNase A). To explain this, the crystal structures of wild-type RNase A and three mutants--F120A, F120G, and F120W--were analyzed up to a 1.4 A resolution. Although the overall backbone structures of all mutant samples were nearly the same as that of wild-type RNase A, except for the C-terminal region of F120G with a high B-factor, two local conformational changes were observed at His119 in the mutants. First, His119 of the wild-type and F120W RNase A adopted an A position, whereas those of F120A and F120G adopted a B position, but the static crystallographic position did not reflect either the efficiency of transphosphorylation or the hydrolysis reaction. Second, His119 imidazole rings of all mutant enzymes were deviated from that of wild-type RNase A, and those of F120W and F120G appeared to be "inside out" compared with that of wild-type RNase A. Only approximately 1 A change in the distance between N(epsilon2) of His12 and N(delta1) of His119 causes a drastic decrease in k(cat), indicating that the active site requires the strict positioning of the catalytic residues. A good correlation between the change in total accessible surface area of the pockets on the surface of the mutant enzymes and enthalpy change in their thermal denaturation also indicates that the effects caused by the replacements are not localized but extend to remote regions of the protein molecule.

PubMed: 11742124
DOI: 10.1110/ps.ps.31102

実験手法

X-RAY DIFFRACTION (1.4 Å)