Purification and crystallization of precursors and autoprocessed enzymes of Flavobacterium glycosylasparaginase: an N-terminal nucleophile hydrolase.
Glycosylasparaginase (GA) represents a novel group of proteins that are activated by self-catalyzed peptide-bond cleavage from a single-chain precursor to yield the two subunits required for hydrolase activity. The wild-type GA precursor autoproteolyzes spontaneously into alpha and beta subunits. Strategies are reported here for purification to homogeneity of GA from Flavobacterium meningosepticum in both single-chain precursor and mature (autoprocessed) forms. The recombinant proteins crystallize in different space groups: P1 and P2(1) for the precursor and mature enzymes, respectively. The precursor crystals diffract to 1.9 A resolution with laboratory X-ray radiation.
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New England Biolabs, 32 Tozer Road, Beverly, MA 01915-5599, USA.
Liao
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Guan
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Guo
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D001227:Aspartylglucosylaminase / Q000737:chemistry* / Q000235:genetics; D001426:Bacterial Proteins / Q000737:chemistry; D003460:Crystallization; D018360:Crystallography, X-Ray; D004789:Enzyme Activation; D005417:Flavobacterium; D016297:Mutagenesis, Site-Directed; D011498:Protein Precursors / Q000737:chemistry*; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics
D001426:Bacterial Proteins; D011498:Protein Precursors; D011994:Recombinant Proteins; D001227:Aspartylglucosylaminase