Membrane‐Bound Stable Glycosyltransferases: Highly Oriented Protein Immobilization by a C‐Terminal Cationic Amphipathic Peptide

Abstract

Gezieltes Verankern von Proteinen: Für ein C‐terminales kationisches amphipathisches Peptid werden spezifische Funktionen in der Bildung von bakteriellen membrangebundenen Glycosyltransferasen nachgewiesen. Der Mechanismus wird in ein allgemeines Konzept übertragen, das die geordnete Immobilisierung von Enzymen auf membranmimetischen Oberflächen ermöglicht, z. B. von rekombinanter α1,3‐Fucosyltransferase aus H. pylori auf magnetischen Kügelchen (siehe Bild).

Research on drug–receptor interactions and prediction of drug activity via oriented immobilized receptor capillary electrophoresis

“…In this case, the active site of biomolecules will be all facing the same direction of the capillary center. Compared to the nonorienting immobilizing methods, oriented immobilization provides biological stability and activity . As an example, Bolivar et al.…”

“…In this case, the active site of biomolecules will be all facing the same direction of the capillary center. Compared to the nonorienting immobilizing methods, oriented immobilization provides biological stability and activity [17][18][19]. As an example, Bolivar et al [17] purified and stabilized a glutamate dehygrogenase from Thermos thermophilus via oriented multisubunit plus multipoint covalent immobilization that permitted to greatly improve the overall enzyme stability.…”

Using open‐tubular capillary electrochromatography with part‐coating column for binding constants determination of β2‐adrenergic receptor with seven drugs

“…Compared to the non-orienting immobilizing methods, oriented immobilization provides receptor or protein with better stability and activity [23][24][25]. The oriented immobilization process of ␤ 2 -AR is same with literature [21].…”

Toward Automated Enzymatic Glycan Synthesis in a Compartmented Flow Microreactor System

“…[3] However, yields are still low or moderate. [8] Key to high enzyme activity is the oriented immobilization on surfaces or polymers mediated by terminal peptides or proteins. Although strict enzyme substrate specificity appears as a disadvantage, a whole enzyme toolbox for glycosylation reactions and synthesis of nucleotide sugars as precursors of glycosyltransferases is now available for enzymatic synthesis of complex glycans.…”

“…Galactokinase (GalK) was combined with UDP-sugar pyrophosphorylase (USP) in an enzyme cascade reaction for the synthesis of UDP-galactose (UDP-Gal, 3) from d-(+)-galactose (Gal,1) via α-d-galactose-1-phosphate (Gal-1-P, 2) in the EM-UDP-Gal. [14] The UDP-GlcA, produced in the EM-UDP-GlcA, was used for glucuronosyltransferase (GlcAT) to transfer the GlcA onto LacNAclinker-tBoc in the EM-GlcAT, resulting in the product non-sulfated human natural killer cell-1 (HNK-1) epitope (8). [13] In the EM-GalT, β1,4-galactosyltransferase (GalT) transfers galactose from UDP-Gal onto a tert-butyloxycarbonyl protected N-acetylglucosamine (GlcNAclinker-tBoc, 6) to synthesize N-Acetyl-d-lactosamine (LacNAc-linker-tBoc, 7).…”