5m6q | pdb_00005m6q

Crystal Structure of Kutzneria albida transglutaminase

spinqualitylabelsall models 5m6q, resolution 1.98Å Show:Asymmetric Unit Biological Assembly Drag the structure with the mouse to rotate   Export Animated Image   Print3D Structural highlights 5m6q is a 2 chain structure with sequence from Kutzneria albida DSM 43870. Full crystallographic information is available from OCA. For a guided tour on the structure components use FirstGlance. Method: X-ray diffraction, Resolution 1.98Å Ligands: CL, CME, MES, PG4 Resources: FirstGlance, OCA, PDBe, RCSB, PDBsum, ProSAT Function W5WHY8_9PSEU Publication Abstract from PubMed Microbial transglutaminases (MTGs) catalyze the formation of Gln-Lys isopeptide bonds and are widely used for the cross-linking of proteins and peptides in food and biotechnological applications (e.g. to improve the texture of protein-rich foods or in generating antibody-drug conjugates). Currently used MTGs have low substrate specificity, impeding their biotechnological use as enzymes that do not cross-react with nontarget substrates (i.e. as bio-orthogonal labeling systems). Here, we report the discovery of an MTG from Kutzneria albida (KalbTG), which exhibited no cross-reactivity with known MTG substrates or commonly used target proteins, such as antibodies. KalbTG was produced in Escherichia coli as soluble and active enzyme in the presence of its natural inhibitor ammonium to prevent potentially toxic cross-linking activity. The crystal structure of KalbTG revealed a conserved core similar to other MTGs but very short surface loops, making it the smallest MTG characterized to date. Ultra-dense peptide array technology involving a pool of 1.4 million unique peptides identified specific recognition motifs for KalbTG in these peptides. We determined that the motifs YRYRQ and RYESK are the best Gln and Lys substrates of KalbTG, respectively. By first reacting a bifunctionalized peptide with the more specific KalbTG and in a second step with the less specific MTG from Streptomyces mobaraensis, a successful bio-orthogonal labeling system was demonstrated. Fusing the KalbTG recognition motif to an antibody allowed for site-specific and ratio-controlled labeling using low label excess. Its site specificity, favorable kinetics, ease of use, and cost-effective production render KalbTG an attractive tool for a broad range of applications, including production of therapeutic antibody-drug conjugates. Discovery of a microbial transglutaminase enabling highly site-specific labeling of proteins.,Steffen W, Ko FC, Patel J, Lyamichev V, Albert TJ, Benz J, Rudolph MG, Bergmann F, Streidl T, Kratzsch P, Boenitz-Dulat M, Oelschlaegel T, Schraeml M J Biol Chem. 2017 Sep 22;292(38):15622-15635. doi: 10.1074/jbc.M117.797811. Epub , 2017 Jul 27. PMID:28751378 From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.

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