Expression, Purification and Crystal Structure of a Truncated Acylpeptide Hydrolase from Aeropyrum pernix K1 (2005)

Content Provider	CiteSeerX
Author	Zheng, Bai-Song Peng, Ying Feng, Yan Lou, Zhi-Yong Zhang, Hai-Feng
Abstract	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-∆(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-∆(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. Key words acylpeptide hydrolase; Aeropyrum pernix K1; crystal structure
File Format	PDF
Publisher Date	2005-01-01
Access Restriction	Open
Subject Keyword	Recombinant Aph Linking Structure Optimal Temperature Truncated Acylpeptide Hydrolase N-terminal Helix Optimum Temperature Thermophilic Archaeon Aeropyrum Pernix K1 Biochemical Experiment Aeropyrum Pernix K1 First Short Helix Truncated Mutant Molecular Replacement N-terminal Hydrolysis Abstract Acylpeptide Hydrolase
Content Type	Text