1C7P

CRYSTAL STRUCTURE OF MUTANT HUMAN LYSOZYME WITH FOUR EXTRA RESIDUES (EAEA) AT THE N-TERMINAL

1C7P の概要

• エントリーDOI: 10.2210/pdb1c7p/pdb
• 関連するPDBエントリー: 1C43 1C45 1C46
• 分子名称: LYSOZYME, SODIUM ION (3 entities in total)
• 機能のキーワード: extra n-terminal residues, hydrolase
• 由来する生物種: Homo sapiens (human)
• 細胞内の位置: Secreted: P61626
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 15190.05

構造登録者

Goda, S.,Takano, K.,Yamagata, Y.,Katakura, Y.,Yutani, K. (登録日: 2000-02-29, 公開日: 2000-04-05, 最終更新日: 2024-11-13)

主引用文献

Goda, S.,Takano, K.,Yamagata, Y.,Katakura, Y.,Yutani, K.
Effect of extra N-terminal residues on the stability and folding of human lysozyme expressed in Pichia pastoris.
Protein Eng., 13:299-307, 2000

PubMed Abstract: A human lysozyme expression system by Pichia pastoris was constructed with the expression vector of pPIC9, which contains the alpha-factor signal peptide known for high secretion efficiency. P. pastoris expressed the human lysozyme at about 300 mg/l broth, but four extra residues (Glu(-4)-Ala(-3)-Glu(-2)-Ala(-1)-) were added at the N-terminus of the expressed protein (EAEA-lysozyme). To determine the effect of the four extra residues on the stability, structures and folding of the protein, calorimetry, X-ray crystal analysis and GuHCl denaturation experiments were performed. The calorimetric studies showed that the EAEA-lysozyme was destabilized by 9.6 kJ/mol at pH 2.7 compared with the wild-type protein, mainly caused by the substantial decrease in the enthalpy change (DeltaH). On the basis of structural information on the EAEA-lysozyme, thermodynamic analyses show that (1) the addition of the four residues slightly affected the conformation in other parts far from the N-terminus, (2) the large decrease in the enthalpy change due to the conformational changes would be almost compensated by the decrease in the entropy change and (3) the decrease in the Gibbs energy change between the EAEA and wild-type human lysozymes could be explained by the summation of each Gibbs energy change contributing to the stabilizing factors concerning the extra residues.

PubMed: 10810162
DOI: 10.1093/protein/13.4.299

実験手法

X-RAY DIFFRACTION (2.4 Å)