What question did this study set out to answer?

To compare wild-type and recombinant tyrocidine synthetase 1 to establish the necessity of posttranslational modification for enzyme activity and nonribosomal peptide biosynthesis.

June 6, 1995Biochemistry

Characterization of tyrocidine synthetase 1 (TY1): Requirement of posttranslational modification for peptide biosynthesis

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Authors

EPEva PfeiferBayreuth Medical Center MPMaja Pavela‐VrančićUniversity of Split HDHans von DoehrenJustus-Liebig-Universität Gießen

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Implication

Comparative biochemical analysis demonstrates that 4'-phosphopantetheine modification is essential for tyrocidine synthetase 1 activity in bacteria, highlighting its role in peptide synthesis.

Key Points

To compare wild-type and recombinant tyrocidine synthetase 1 to establish the necessity of posttranslational modification for enzyme activity and nonribosomal peptide biosynthesis.
Compared wild-type TY1 isolated from Bacillus brevis ATCC 8185 with recombinant TY1 heterologously expressed in Escherichia coli.
Analyzed intact protein mass and tryptic fragments using matrix-assisted laser desorption mass spectrometry (MALDI) to evaluate cofactor attachment at Ser562.
Measured amino acid-dependent ATP-PPi exchange specificity, reverse adenylation kinetics, and aminoacylation efficiency between enzyme forms.
MALDI analysis revealed recombinant TY1 had a molecular mass of 122,516 ± 120 Da and lacked the 4'-phosphopantetheine cofactor at the putative Ser562 binding site.
Recombinant TY1 demonstrated a reverse adenylation rate of only 22% relative to the wild-type enzyme.
Aminoacylation dropped from approximately 60% in wild-type TY1 to 1.4% in the overexpressed recombinant protein, while substrate activation profiles remained identical.

Cite This Study

Pfeifer et al. (1995) studied this question.

synapsesocial.com/papers/6a8beb18c12eaf65c8b2240f https://doi.org/10.1021/bi00022a019

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Implication

Comparative biochemical analysis demonstrates that 4'-phosphopantetheine modification is essential for tyrocidine synthetase 1 activity in bacteria, highlighting its role in peptide synthesis.

Characterization of tyrocidine synthetase 1 (TY1): Requirement of posttranslational modification for peptide biosynthesis

Discussion

Member takes

Implication

Key Points

Cite This Study

Characterization of tyrocidine synthetase 1 (TY1): Requirement of posttranslational modification for peptide biosynthesis

Discussion

Member takes

Implication

Key Points

Cite This Study

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