Characterization of extracellular proteases from Trametes trogii

Caporale, Carlo; Garzillo, Anna Maria Vittoria; Caruso, Carla; Buonocore, Vincenzo

doi:10.1016/0031-9422(96)83284-5

Cited by 19 publications

(13 citation statements)

References 24 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Coprinus cinereus produces a leucine aminopeptidase in meiotic prophase tissue [18], Lyophyllum cinerascens and Grifola frondosa aminopeptidases have been purified and biochemically characterized [19,20], and the mycelial Trametes trogi aminopeptidase has been detected in culture supernatants [21]. Coprinus cinereus produces a leucine aminopeptidase in meiotic prophase tissue [18], Lyophyllum cinerascens and Grifola frondosa aminopeptidases have been purified and biochemically characterized [19,20], and the mycelial Trametes trogi aminopeptidase has been detected in culture supernatants [21].…”

Section: Introductionmentioning

confidence: 99%

“…Production and roles of the aminopeptidases in basidiomycete fungi have been studied in a few systems. Coprinus cinereus produces a leucine aminopeptidase in meiotic prophase tissue [18], Lyophyllum cinerascens and Grifola frondosa aminopeptidases have been purified and biochemically characterized [19,20], and the mycelial Trametes trogi aminopeptidase has been detected in culture supernatants [21].…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Purification and characterization of aminopeptidase (pumAPE) from Ustilago maydis

Mercado-Flores

Noriega-Reyes

Ramı́rez-Zavala

et al. 2004

FEMS Microbiology Letters

View full text Add to dashboard Cite

The aminopeptidase pumAPE was purified from the haploid fungus Ustilago maydis FB1 strain. The purification procedure consisted of ammonium sulfate fractionation and three chromatographic steps, which included anion-exchange, hydrophobic interaction, and gel filtration chromatography, resulting in a 23% recovery. The molecular mass of the dimeric enzyme was estimated to be 110 kDa and 58 kDa by gel filtration chromatography and SDS-PAGE, respectively. Enzymatic activity was optimal at pH 7.0 and at 35 degrees C toward Lys-pNA and the pI was determined to be 5.1. The enzyme was inhibited by EDTA-Na2, 1,10- phenanthroline, bestantin, PMSF and several divalent cations (Cu2+, Hg2+ and Zn2+). The aminopeptidase showed a preference for lysine and arginine in the N-position. The K(m) value was 54.4 microM and the Vmax value was 408 micromolmin(-1)mg(-1) for Lys-pNA.

show abstract

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Purification and characterization of aminopeptidase (pumAPE) from Ustilago maydis

Mercado-Flores

Noriega-Reyes

Ramı́rez-Zavala

et al. 2004

FEMS Microbiology Letters

View full text Add to dashboard Cite

show abstract

“…These difficulties have been greatly reduced using modern pulsed-liquid phase sequencers, since the carryover is a well quantifiable phenomenon and, despite the yield problems, the identification of several amino acid residues at each step requires just a little care and experience. [43][44][45][46][47][48][49][50][51][52][53][54] Thus, digesting a protein of known sequence by any agent, it is possible to assess the sequence of each fragment present in the mixture and evaluate its amount. Although the absolute quantification is underestimated due to the initial coupling yield of Edman degradation (60-65%), the data can be compared and supply quantitative information since they suffer for a similar error.…”

Section: Results and Discussion Protein Digestions Hplc And Sequencmentioning

confidence: 99%

“…Different strategies in assessing protein sequences have been reported [43][44][45][46][47][48][49] together with methods for determining the hydrolysis pathway of polypeptides by unknown digesting agents. [50][51][52][53] These approaches, which have been recently reviewed, 54 do not require any purification step and are based upon the sequence determination of the fragments in mixture; moreover, sequence data being quantitative, the amount of each fragment can be evaluated. [43][44][45][46][47][48][49][50][51][52][53][54] The method can also be used to get rapid information on the tertiary structure of proteins in solution.…”

Section: Introductionmentioning

confidence: 99%

Probing the modelled structure of Wheatwin1 by controlled proteolysis and sequence analysis of unfractionated digestion mixtures

et al. 1999

Self Cite

View full text Add to dashboard Cite

We set up a method to get rapid information on the three-dimensional structure of peptide and proteins of known sequence. Both native and alkylated polypeptide is hydrolyzed with a number of proteases at different digestion times and the resulting mixtures are compared by HPLC analysis to establish the differences in the hydrolysis pathways of the folded and unfolded molecule. Then, the unfractionated digestion mixtures of the native polypeptide are submitted to automatic sequence analysis to identify the hydrolysis sites. The sequence of each fragment present in the mixtures is reconstructed and its amount determined by quantitative data of the sequence analyses. We used this approach to determine the amino acid surface accessibility of wheatwin1, a pathogenesis-related protein from wheat, and constructed a predictive three-dimensional model based on the knowledge of the tertiary structure of barwin, a highly homologous protein from barley. The procedure allowed us to quickly identify and quantify the hydrolysis at the susceptible bonds which could be classified as exposed, partially hidden, or inaccessible. The results were useful to evidentiate and discuss concordances and differences between experimental and model predicted accessibilities of amino acid residues. Proteins 1999;36:192-204.

show abstract

“…The most critical points in identifying the residues present at each step of sequence analysis of peptide mixtures are due to the carryover of the Edman degradation and to the partial destruction of labile residues (e.g., Ser and Thr). In recent years, the use of modern pulsed-liquid phase sequencers equipped on-line with PTH analyzers has greatly simplified these problems Caporale et al, 1993Caporale et al, , 1994Caporale et al, , 1996aPetrilli et al, 1994). In fact, the identification of a number of residues at each step needs just a little of experience, since the carryover is a phenomenon that can be quantified and labile PTH residues are easily identified (e.g., Set and Thr are simultaneously identified both as PTH-amino acids and PTH-A-amino acids).…”

Section: Sequence Datamentioning

confidence: 99%

Determination of the primary structure of homologous proteins by sequence analysis of peptide mixtures

1996

Self Cite

View full text Add to dashboard Cite

We propose a rapid method to determine the primary structure of a protein knowing the sequence of a homologous protein. The method consists in submitting both the reduced and alkylated proteins to an enzymatic or chemical hydrolysis and performing the sequence analysis of the peptide mixtures. The assessment of the unknown sequence and the degree of identify of the two proteins are reached by comparing the two sequence analyses. The sequences of all the possible peptides present in the two mixtures are reconstructed and the differences in the two sequences are determined. If necessary, the differences can be confirmed by performing a mass spectrometric analysis of the two mixtures. We used this procedure on two homologous proteins of known sequence to furnish an application example of the method.

show abstract

Characterization of extracellular proteases from Trametes trogii

Cited by 19 publications

References 24 publications

Purification and characterization of aminopeptidase (pumAPE) from Ustilago maydis

Purification and characterization of aminopeptidase (pumAPE) from Ustilago maydis

Probing the modelled structure of Wheatwin1 by controlled proteolysis and sequence analysis of unfractionated digestion mixtures

Determination of the primary structure of homologous proteins by sequence analysis of peptide mixtures

Contact Info

Product

Resources

About