Streptomyces griseus aminopeptidase: X-ray crystallographic structure at 1.75 Å resolution

Greenblatt, H.M.; Almog, Orna; Maras, Bruno; Spungin-Bialik, A.; Barra, Donatella; Blumberg, Shmaryahu; Shoham, G.

doi:10.1006/jmbi.1996.0729

Cited by 119 publications

(149 citation statements)

References 34 publications

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“…In S. griseus and Aeromonas proteolytica secreted aminopeptidases, two residues, His and Asp, bind a first Zn 2+ ion, two additional residues His and Glu bind a second Zn 2+ ion, while a second Asp residue bridges the (Hall, 1999) using PAM250 similarity matrix. two Zn 2+ ions (Greenblatt et al, 1997;Hasselgren et al, 2001). Substitution of Zn 2+ by different divalent ions in S. griseus secreted aminopeptidase is affected by Ca 2+ , and has variable effects (Ben-Meir et al, 1993;Lin et al, 1997;Hasselgren et al, 2001).…”

Section: Discussionmentioning

confidence: 99%

Aminopeptidases and dipeptidyl-peptidases secreted by the dermatophyte Trichophyton rubrum

et al. 2005

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The nature of secreted aminopeptidases in Trichophyton rubrum was investigated by using a reverse genetic approach. T. rubrum genomic and cDNA libraries were screened with Aspergillus spp. and Saccharomyces cerevisiae aminopeptidase genes as the probes. Two leucine aminopeptidases, ruLap1 and ruLap2, and two dipeptidyl-peptidases, ruDppIV and ruDppV, were characterized and compared to orthologues secreted by Aspergillus fumigatus using a recombinant protein from Pichia pastoris. RuLap1 is a 33 kDa nonglycosylated protein, while ruLap2 is a 58-65 kDa glycoprotein. The hydrolytic activity of ruLap1, ruLap2 and A. fumigatus orthologues showed various preferences for different aminoacyl-7-amido-4-methylcoumarin substrates, and various sensitivities to inhibitors and cations. ruDppIV and ruDppV showed similar activities to A. fumigatus orthologues. In addition to endopeptidases, the four aminopeptidases ruLap1, ruLap2, ruDppIV and ruDppV were produced by T. rubrum in a medium containing keratin as the sole nitrogen source. Synergism between endo-and exopeptidases is likely to be essential for dermatophyte virulence, since these fungi grow only in keratinized tissues.

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Section: Discussionmentioning

confidence: 99%

Aminopeptidases and dipeptidyl-peptidases secreted by the dermatophyte Trichophyton rubrum

et al. 2005

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“…The substrate specificity of VpAP and SgAP is similar, both cleaving preferentially hydrophobic amino acids especially leucine occupying the N-terminal position of proteins or peptides (25,28,29). Although the amino acid sequences of VpAP and SgAP show relatively low homology (ϳ29% identity) their three-dimensional structures are almost superimposable (30). Both contain two adjacent zinc atoms in their active sites that are required for activity, and the five amino acid residues involved in coordination of the two active site zinc atoms (two His, two Asp, and one Glu) are identical in the two enzymes and overlap closely (30).…”

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confidence: 99%

“…Although the amino acid sequences of VpAP and SgAP show relatively low homology (ϳ29% identity) their three-dimensional structures are almost superimposable (30). Both contain two adjacent zinc atoms in their active sites that are required for activity, and the five amino acid residues involved in coordination of the two active site zinc atoms (two His, two Asp, and one Glu) are identical in the two enzymes and overlap closely (30). The gene coding for VpAP (but not SgAP) has been cloned (31,32).…”

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confidence: 99%

A Secreted Aminopeptidase of Pseudomonas aeruginosa

Cahan¹,

Axelrad²,

Safrin³

et al. 2001

Journal of Biological Chemistry

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Using leucine-p-nitroanilide (Leu-pNA) as a substrate, we demonstrated aminopeptidase activity in the culture filtrates of several Pseudomonas aeruginosa strains. The aminopeptidase was partially purified by DEAE-cellulose chromatography and found to be heat stable. The apparent molecular mass of the enzyme was ϳ56 kDa; hence, it was designated AP 56 . Heating (70°C) of the partially purified aminopeptidase preparations led to the conversion of AP 56 to a ϳ28-kDa protein (AP 28 ) that retained enzyme activity, a reaction that depended on elastase (LasB). The pH optimum for Leu-pNA hydrolysis by AP 28 was 8.5. This activity was inhibited by Zn chelators but not by inhibitors of serine-or thiol-proteases, suggesting that AP 28 is a Zn-dependent enzyme. Of several amino acid p-nitroanilide derivatives examined, Leu-pNA was the preferred substrate. The sequences of the first 20 residues of AP 56 and AP 28 were determined. A search of the P. aeruginosa genomic data base revealed a perfect match of these sequences with positions 39 -58 and 273-291, respectively, in a 536-amino acid residue open reading frame predicted to encode an aminopeptidase. A search for sequence similarities with other proteins revealed 52% identity with Streptomyces griseus aminopeptidase, ϳ35% identity with Saccharomyces cerevisiae aminopeptidase Y and a hypothetical aminopeptidase from Bacillus subtilis, and 29 -32% with Aeromonas caviae, Vibrio proteolyticus, and Vibrio cholerae aminopeptidases. The residues potentially involved in zinc coordination were conserved in all these proteins. Thus, P. aeruginosa aminopeptidase may belong to the same family (M28) of metalloproteases.

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“…Similarly to the transferrin receptor, GCPII probably exists as a homodimer under physiological conditions and the dimerization seems to be essential for its hydrolytic activity [15]. The protein is proposed to consist of six domains: the N-terminal cytoplasmic tail (amino acids [1][2][3][4][5][6][7][8][9][10][11][12][13][14][15][16][17][18], the helical transmembrane region (amino acids , and four extracellular domains spanning amino acids 44-150 (domain C), 151-274 (domain D), 275-586 (domain E), and 587-750 (domain F). While the domain spanning amino acids 275-586 is believed to be the catalytic domain, the importance/ function of the three remaining extracellular domains is unknown [16].…”

mentioning

confidence: 99%

“…While the domain spanning amino acids 275-586 is believed to be the catalytic domain, the importance/ function of the three remaining extracellular domains is unknown [16]. The putative catalytic domain of GCPII is homologous to aminopeptidases from S. griseus and V. proteolyticus whose crystal structures have been solved at 1.75 Å and 1.8 Å resolution, respectively [17,18]. By analogy with the Vibrio aminopeptidase and the alignment of partial amino acid sequences from human GCPII, human transferrin receptor, yeast aminopeptidase Y, S. griseus aminopeptidase, and Caenorhabditis elegans mGCP fragment, His377, Asp387, Glu425, Asp453 and His553 were proposed to be the zinc ligands of GCPII [16].…”

mentioning

confidence: 99%

Amino acids at the N‐ and C‐termini of human glutamate carboxypeptidase II are required for enzymatic activity and proper folding

Bařinka

Mlčochová

Šácha

et al. 2004

European Journal of Biochemistry

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Human glutamate carboxypeptidase II (GCPII) is a co-catalytic metallopeptidase and its putative catalytic domain is homologous to the aminopeptidases from Vibrio proteolyticus and Streptomyces griseus. In humans, the enzyme is expressed predominantly in the nervous system and the prostate. The prostate form, termed prostate-specific membrane antigen, is overexpressed in prostate cancer and is used as a diagnostic marker of the disease. Inhibition of the form of GCPII expressed in the central nervous system has been shown to protect against ischemic injury in experimental animal models. Human GCPII consists of 750 amino acids, and six individual domains were predicted to constitute the protein structure. Here, we report the analysis of the contribution of these putative domains to the structure/ function of recombinant human GCPII. We cloned 13 mutants of human GCPII that are truncated or extended at one or both the N-and C-termini of the GCPII sequence.The clones were used to generate stably transfected Drosophila Schneider's cells, and the expression and carboxypeptidase activities of the individual protein products were determined. The extreme C-terminal region of human GCPII was found to be critical for the hydrolytic activity of the enzyme. The deletion of as few as 15 amino acids from the C-terminus was shown to completely abolish the enzymatic activity of GCPII. Furthermore, the GCPII carboxypeptidase activity was abrogated upon removal of more than 60 amino acid residues from the N-terminus of the protein. Overall, these results clearly show that amino acid segments at the N-and C-termini of the ectodomain of GCPII are essential for its carboxypeptidase activity and/or proper folding.

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Streptomyces griseus aminopeptidase: X-ray crystallographic structure at 1.75 Å resolution

Cited by 119 publications

References 34 publications

Aminopeptidases and dipeptidyl-peptidases secreted by the dermatophyte Trichophyton rubrum

Aminopeptidases and dipeptidyl-peptidases secreted by the dermatophyte Trichophyton rubrum

A Secreted Aminopeptidase of Pseudomonas aeruginosa

Amino acids at the N‐ and C‐termini of human glutamate carboxypeptidase II are required for enzymatic activity and proper folding

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