Pepsin‐catalyzed peptide synthesis

Pellegrini, Antonio; Luigilusis, Pier

doi:10.1002/bip.1978.360171105

Cited by 25 publications

(6 citation statements)

References 20 publications

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“…Peptide synthesis by the reverse proteolytic reaction is well-known. Detailed papers and extensive review articles have been presented in the past by Jakubke [17] and by other authors [18] [19], and, within the field of the origin of life, scenarios of alternate dry and wet environment have been proposed as conditions for bond-formation [12].…”

mentioning

confidence: 99%

The Production ofde novo Folded Proteins by a Stepwise Chain Elongation: A Model for Prebiotic Chemical Evolution of Macromolecular Sequences

Chessari

Thomas

Polticelli

et al. 2006

C&B

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We describe an experimental procedure to mimic the formation of long (over 40 residues) co-oligopetide sequences in many identical copies which may have occurred in the prebiotic molecular evolution. The basic hypothesis is that chain formation is based on the stepwise fragment condensation of randomly generated short oligopeptides, whereby the elongation takes place under the contingent environmental constraints (solubility, pH, salinity), which eliminate most of the products, and thus determine the selection towards one particular small set of chains. The present work aims at verifying the validity of this scheme. In order to do so, we utilize a classic synthetic procedure based on the Merrifield solid-phase synthesis of peptides for the synthesis of randomly produced peptides as well as for their stepwise fragment condensation. Thus, starting from a library of peptides with n=10, the first condensation step produces a library of 16 peptides with 20 residues each (n=20), of which only four remain water-soluble and, therefore, capable to undergo the next fragment condensation step. This gives rise to 16 peptides with n=30, out of which twelve precipitate out under the chosen pH and buffer conditions and are eliminated. Finally, a 44-residue-long water-soluble de novo protein is obtained. This has no homologies or similarities with extant proteins, and, based on circular dichroism (CD), it assumes a stable three-dimensional folding. In agreement with CD data, molecular-modelling simulations suggest an helical fold for the protein with poor, if any, structural homology with known proteins. The implication of this procedure as a general mechanism for the etiology of de novo macromolecular sequences and globular proteins in the origin of life is briefly discussed.

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mentioning

confidence: 99%

The Production ofde novo Folded Proteins by a Stepwise Chain Elongation: A Model for Prebiotic Chemical Evolution of Macromolecular Sequences

Chessari

Thomas

Polticelli

et al. 2006

C&B

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“…However, the principle of microscopic reversibility [20] prescribes that specific base catalysis, the reverse of specific acid catalysis, .ought to facilitate peptide synthesis if the forward reaction is catalysed by oxonium ions. The reaction of hydroxide ions, however, could hardly be important at acidic pH, where peptide synthesis by pepsin takes place [21]. The other alternative of the leaving group protonation involves proton transfer from the carbonyl oxygen by the participation of Asp 215 [18].…”

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confidence: 99%

The mechanism of action of aspartic proteases involves ‘push‐pull’ catalysis

Polgár

1987

FEBS Letters

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In accord with the available kinetic and X-ray crystallographic data, it is proposed that the two catalytically competent carboxyl groups of aspartic proteases constitute a functional unit which mediates the proton from the attacking water molecule to the leaving nitrogen atom of the substrate. Protonation of this nitrogen atom has been the main issue of the previous mechanistic proposals. The first step of the present mechanism involves proton transfer from the water to the aspartic diad and concurrently another proton transfer from the diad to the carbonyl oxygen of the scissile peptide bond. These proton transfers provide the driving force for the bond formation between the substrate and water, which leads to the formation of a tetrahedral intermediate. The intermediate breaks down to products by a similar facilitation, i.e. by concerted general acid-base catalysis, which involves simultaneous proton transfers from the intermediate to the diad and from the diad to the leaving nitrogen of the substrate. The symmetrical mechanism of the formation and decomposition of the tetrahedral adduct resembles that found in the serine protease catalysis.Pepsin; Aspartic protease; Enzyme mechanism Aspartic proteases include several important enzymes, such as pepsin, chymosin, renin, cathepsin D and the proteases isolated from numerous fungi. The amino acid sequences of all these enzymes are homologous, in particular around the active-site residues. Comprehensive reviews on aspartic proteases have been published [1][2][3]. The threedimensional structure of pepsin [4] and three microbial aspartic proteases including penicillopepsin [5,6], Rhizopus chinensin protease [7] and Endothiaparasitica protease [7] has been reported. The tertiary structures of pepsin and the microbial Correspondence address: L. Polg~ir,

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“…The results obtained thus far in our laboratory, and by other investigators (38,43), indicate that the aspartyl proteinases may be useful for coupling reactions in which it is desired to join two hydrophobic amino acid residues. An example is the pepsin-catalyzed condensation, recently performed by Dr. MICHAEL GELB in my laboratory, of Z-Tyr(Bzl)-Gly-Gly-Phe-OH with H-Met-Arg(NO2)-Phe-OBzl to produce fully-protected pro-Met-enkephalin.…”

Section: Pepsin-catalyzed Peptide Bondmentioning

confidence: 54%

Pepsin and thermolysin as catalysts of peptide bond synthesis

Fruton

1984

Carlsberg Res. Commun.

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The early studies on proteinase-catalyzed peptide bond synthesis are summarized as background for a report on some recent work in our laboratory on pepsin and thermolysin. The primary and secondary specificity of these two enzymes have been found to be suitable for the preparative synthesis of oligopeptides by the condensation of two hydrophobic L-amino acid residues. Kinetic data are presented showing that the rate of entry of each of the two components in a thermolysin-catalyzed condensation reaction depends on the nature of the other component, and that this effect is related to differences in the transition-state conformation of the enzyme-reactant complexes.

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Pepsin‐catalyzed peptide synthesis

Cited by 25 publications

References 20 publications

The Production ofde novo Folded Proteins by a Stepwise Chain Elongation: A Model for Prebiotic Chemical Evolution of Macromolecular Sequences

The Production ofde novo Folded Proteins by a Stepwise Chain Elongation: A Model for Prebiotic Chemical Evolution of Macromolecular Sequences

The mechanism of action of aspartic proteases involves ‘push‐pull’ catalysis

Pepsin and thermolysin as catalysts of peptide bond synthesis

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