Expression, Purification and Crystal Structure of a Truncated Acylpeptide Hydrolase from Aeropyrum pernix K1

  1. Hai-Feng ZHANG2,
  2. Bai-Song ZHENG1,
  3. Ying PENG2,
  4. Zhi-Yong LOU2,
  5. Yan FENG1,*,
  6. Zi-He RAO2,*

Article first published online: 27 SEP 2005

DOI: 10.1111/j.1745-7270.2005.00085.x

Issue

Acta Biochimica et Biophysica Sinica

Acta Biochimica et Biophysica Sinica

Volume 37, Issue 9, pages 613–617, September 2005

Additional Information

How to Cite

ZHANG, H.-F., ZHENG, B.-S., PENG, Y., LOU, Z.-Y., FENG, Y. and RAO, Z.-H. (2005), Expression, Purification and Crystal Structure of a Truncated Acylpeptide Hydrolase from Aeropyrum pernix K1. Acta Biochimica et Biophysica Sinica, 37: 613–617. doi: 10.1111/j.1745-7270.2005.00085.x

Author Information

  1. 1

    Key Laboratory for Molecular Enzymology and Engineering of Ministry of Education, Jilin University, Changchun 130023, China;

  2. 2

    Laboratory of Structural Biology, Department of Biological Sciences and Biotechnology and Protein Sciences Laboratory of Ministry of Education, Tsinghua University, Beijing 100084, China

* *Corresponding authors: Yan FENG: Tel, 86–431–8987975; E-mail, yfeng@jlu.edu.cn Zi-He RAO: Tel, 86–10–62771493; Fax, 86–10–62773145; E-mail, raozh@xtal.tsinghua.edu.cn

Publication History

  1. Issue published online: 27 SEP 2005
  2. Article first published online: 27 SEP 2005
  3. Received: March 20, 2005 Accepted: June 7, 2005

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Keywords:

  • acylpeptide hydrolase;
  • Aeropyrum pernix K1;
  • crystal structure

Abstract Acylpeptide hydrolase (APH) catalyzes the N-terminal hydrolysis of Nα-acylpeptides to release Nα-acylated amino acids. The crystal structure of recombinant APH from the thermophilic archaeon Aeropyrum pernix K1 (apAPH) was reported recently to be at a resolution of 2.1 Å using X-ray diffraction. A truncated mutant of apAPH that lacks the first short α-helix at the N-terminal, apAPH-α(1–21), was cloned, expressed, characterized and crystallized. Data from biochemical experiments indicate that the optimum temperature of apAPH is decreased by 15 °C with the deletion of the N-terminal α-helix. However, the enzyme activity at the optimal temperature does not change. It suggests that this N-terminal α-helix is essential for thermostability. Here, the crystal structure of apAPH-α(1–21) has been determined by molecular replacement to 2.5 å. A comparison between the two structures suggests a difference in thermostability, and it can be concluded that by adding or deleting a linking structure (located over different domains), the stability or even the activity of an enzyme can be modified.

Edited by Wei-Min GONG

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