Structural and biochemical characterization of a novel aminopeptidase from human intestine.

The Journal of Biological Chemistry
Jan TykvartJan Konvalinka

Abstract

N-acetylated α-linked acidic dipeptidase-like protein (NAALADase L), encoded by the NAALADL1 gene, is a close homolog of glutamate carboxypeptidase II, a metallopeptidase that has been intensively studied as a target for imaging and therapy of solid malignancies and neuropathologies. However, neither the physiological functions nor structural features of NAALADase L are known at present. Here, we report a thorough characterization of the protein product of the human NAALADL1 gene, including heterologous overexpression and purification, structural and biochemical characterization, and analysis of its expression profile. By solving the NAALADase L x-ray structure, we provide the first experimental evidence that it is a zinc-dependent metallopeptidase with a catalytic mechanism similar to that of glutamate carboxypeptidase II yet distinct substrate specificity. A proteome-based assay revealed that the NAALADL1 gene product possesses previously unrecognized aminopeptidase activity but no carboxy- or endopeptidase activity. These findings were corroborated by site-directed mutagenesis and identification of bestatin as a potent inhibitor of the enzyme. Analysis of NAALADL1 gene expression at both the mRNA and protein levels revealed ...Continue Reading

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Citations

Mar 31, 2015·Virology·Jan KonvalinkaBarbara Müller
Jan 13, 2018·Proteomics·Antonio PalombaSergio Uzzau
Sep 1, 2018·Protein Science : a Publication of the Protein Society·Jakub PtacekCyril Barinka
Nov 21, 2015·Journal of Nuclear Medicine : Official Publication, Society of Nuclear Medicine·Thomas PykaMatthias Eiber

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