Taxonomic distribution of proteins containing Aspzincin_M35 domains

The tree below includes only several representative species and genera. The complete taxonomic breakdown of all proteins containing Aspzincin_M35 domains can be accessed here. Click the counts or percentage values to display the corresponding proteins.

A quick solution: ab initio structure determination of a 19 kDa metalloproteinaseusing ACORN.

McAuley KE, Jia-Xing Y, Dodson EJ, Lehmbeck J, Ostergaard PR, Wilson KS
Acta Crystallogr D Biol Crystallogr. 2001; 57: 1571-8

A data set from the metalloproteinase deuterolysin was collected at atomicresolution (1.0 A) with synchrotron radiation. The high resolution allowed thestructure to be solved with the new direct-methods program ACORN using thecoordinates of the Zn atom as a starting point. The phases obtained from ACORNwere of sufficient quality to allow automated building to be carried out inARP/wARP. Minimal manual rebuilding of the model was required and the structuredetermination was completed using the maximum-likelihood refinement programREFMAC. The whole process, starting from the processed and merged data and endingwith a refined model, required less than 6 h of computational time.

Kinetic characterization of lysine-specific metalloendopeptidases from Grifolafrondosa and Pleurotus ostreatus fruiting bodies.

Nonaka T, Hashimoto Y, Takio K
J Biochem. 1998; 124: 157-62

Two zinc-metalloendopeptidases, GFMEP (accession number P81054) and POMEP(accession number P81055), from the fruiting bodies of two edible mushrooms,Grifola frondosa and Pleurotus ostreatus, respectively, specifically hydrolyzepeptidyl-lysine bonds (-X-Lys-) in polypeptides. To understand detailed substratespecificities and kinetic characters of these enzymes, we have synthesizedvarious intramolecularly quenched fluorescent peptide substrates and determinedtheir kinetic constants with these substrates. Each synthesized fluorogenicpeptide has a fluorescent residue, tryptophan, at its carboxyl terminus and aquenching group, dinitrophenyl (Dnp), at its amino terminus. Quenching of the Trpfluorescence in an intact substrate is relieved on hydrolysis of the -X-Lys-bond, giving rise to a continuous increase in fluorescence. The octapeptidesubstrate, Dnp-Ser-Thr-Ala-Thr-Lys-Leu-Ser-Trp, was an efficient substrate forboth enzymes, the kcat/Km values being 9.8 x 10(6) and 7.0 x 10(5) M-1.s-1 forGF- and POMEP, respectively. Peptides with aspartic acid adjacent to the Lysresidue were found to be poor substrates for both enzymes. Neither the shortestpeptide, Dnp-Thr-Lys-Trp, nor peptides with substitution of L-Arg, L-ornithine,or D-Lys for Lys were hydrolyzed by either enzyme. These results confirmed thestrict specificities of GF- and POMEP toward the peptide bond, -X-Lys-.Substitution of Co2+ for Zn2+ enhanced the activity, while the Km values werecomparable. All peptides not hydrolyzed by either enzyme had inhibitory effectson GFMEP activity. The active site structure is discussed in relation to theseobservations.