Related Experiment Video

Updated: Mar 16, 2026

Monitoring the Assembly of a Secreted Bacterial Virulence Factor Using Site-specific Crosslinking
11:33

Monitoring the Assembly of a Secreted Bacterial Virulence Factor Using Site-specific Crosslinking

Published on: December 17, 2013

6.6K

Structure of the Dispase Autolysis-inducing Protein from Streptomyces mobaraensis and Glutamine Cross-linking Sites

David Fiebig1, Stefan Schmelz2, Stephan Zindel1

  • 1Department of Chemical Engineering and Biotechnology, University of Applied Sciences of Darmstadt, 64287 Darmstadt, Germany, and the Department of Chemistry and.

The Journal of Biological Chemistry
|August 6, 2016
PubMed

Insights

Transglutaminase from Streptomyces mobaraensis (MTG) cross-links proteins. This study identified specific glutamine sites on dispase autolysis-inducing protein (DAIP), revealing MTG

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Structural Biology

Background:

  • Transglutaminase from Streptomyces mobaraensis (MTG) is crucial for protein cross-linking.
  • Dispase autolysis-inducing protein (DAIP) is an intrinsic substrate of MTG.
  • Understanding MTG's substrate specificity is key to its applications.

Purpose of the Study:

  • To determine the structure of DAIP.
  • To identify the specific glutamine residues on DAIP targeted by MTG.
  • To elucidate the determinants of MTG's modification specificity and efficiency.

Main Methods:

  • Recombinant DAIP production in E. coli.
  • Site-directed mutagenesis to create DAIP variants.
  • Biochemical assays using biotin cadaverine incorporation to map cross-linking sites.
  • Structural analysis of DAIP.

Main Results:

  • Identified and ranked the preference of MTG for specific glutamine residues in DAIP (Gln-39 ≫ Gln-298 > Gln-345 ∼ Gln-65 ≫ Gln-144).
  • Structural data revealed that preferred glutamines cluster on the DAIP β-propeller structure.
  • Proposed a model for targeted DAIP cross-linking by MTG following self-assembly.

Conclusions:

  • The study provides the first structural and biochemical characterization of a physiological MTG substrate.
  • DAIP's glutamine residues exhibit differential accessibility and reactivity towards MTG.
  • Findings offer novel insights into MTG specificity and efficiency, relevant for protein modification applications.
Keywords:
Dispase autolysis inducing proteinStreptomyces mobaraensisX-ray crystallographyenzymeglutamine cross-linking sitesmicrobial transglutaminaseprotein chemical modificationprotein chemistrytertiary structure

More Related Videos

Purification of Hsp104, a Protein Disaggregase07:17

Purification of Hsp104, a Protein Disaggregase

Published on: September 30, 2011

17.8K
Exploring Caspase Mutations and Post-Translational Modification by Molecular Modeling Approaches05:56

Exploring Caspase Mutations and Post-Translational Modification by Molecular Modeling Approaches

Published on: October 13, 2022

1.9K

Related Concept Videos

Caspases01:24

Caspases

Caspase, a family of cysteine proteases, serve as effectors in apoptosis. The ced3 gene in C.elegans was first identified to be involved in apoptosis. This gene encodes the ced-3 caspase that is similar to the interleukin-1-beta converting enzyme or ICE in mammals. In addition to apoptosis, caspases also function in the inflammatory response. Inflammatory caspases are essential in activating pro-inflammatory cytokines that recruit immune cells and block the replication of pathogens inside...
14.5K
Tagging and Fusion Proteins01:24

Tagging and Fusion Proteins

Proteins are involved in several cellular processes and biochemical reactions. Analyzing a specific protein of interest requires it to be isolated from the other proteins in the cell. This is achieved by overexpressing the specific gene in a suitable host to produce large quantities of the target protein. A tag or label is recombined with the gene to produce a fusion protein containing the target protein and the tag. The tags on these fusion proteins can then be used for easy detection and...
8.7K
Peptidoglycan Synthesis01:28

Peptidoglycan Synthesis

Structure of PeptidoglycanPeptidoglycan is a vital structural component of the bacterial cell wall, providing mechanical strength and shape to the cell. It consists of repeating units of two sugars—N-acetylglucosamine (NAG) and N-acetylmuramic acid (NAM)—linked by β-1,4 glycosidic bonds. These sugar chains are cross-linked by short peptide chains, forming a mesh-like polymer that surrounds the bacterial plasma membrane.Cytoplasmic Phase – Precursor SynthesisPeptidoglycan...
3.8K