1P4K

CRYSTAL STRUCTURE OF THE GLYCOSYLASPARAGINASE PRECURSOR D151N MUTANT

Summary for 1P4K

• Entry DOI: 10.2210/pdb1p4k/pdb
• Related: 1P4V
• Descriptor: N(4)-(Beta-N-acetylglucosaminyl)-L-asparaginase, GLYCEROL (2 entities in total)
• Functional Keywords: alpha beta, beta alpha, sandwich, hydrolase
• Biological source: Elizabethkingia meningoseptica
• Cellular location: Periplasm: Q47898
• Total number of polymer chains: 2
• Total formula weight: 64725.67

Authors

Qian, X.,Guan, C.,Guo, H.C. (deposition date: 2003-04-23, release date: 2003-05-06, Last modification date: 2024-02-14)

Primary citation

Qian, X.,Guan, C.,Guo, H.C.
A dual role for an aspartic acid in glycosylasparaginase autoproteolysis.
Structure, 11:997-1003, 2003

PubMed Abstract: Glycosylasparaginase uses an autoproteolytic processing mechanism, through an N-O acyl shift, to generate a mature/active enzyme from a single-chain precursor. Structures of glycosylasparaginase precursors in complex with a glycine inhibitor have revealed the backbone in the immediate vicinity of the scissile peptide bond to be in a distorted trans conformation, which is believed to be the driving force for the N-O acyl shift to break the peptide bond. Here we report the effects of point mutation D151N. In addition to the loss of the base essential in autoproteolysis, this mutation also eradicates the backbone distortion near the scissile peptide bond. Binding of the glycine inhibitor to the autoproteolytic site of the D151N mutant does not restore the backbone distortion. Therefore, Asp151 plays a dual role, acting as the general base to activate the nucleophile and holding the distorted trans conformation that is critical for initiating an N-O acyl shift.

PubMed: 12906830
DOI: 10.1016/S0969-2126(03)00150-3

Experimental method

X-RAY DIFFRACTION (1.9 Å)