Biotin acceptor peptide

Using the genetic approach the biotinylated obelin was obtained. A recombinant apoobelin capable of being biotinylated in vivo in E. coli cells with BirA was constructed by fusing in-frame a synthetic DNA-fragment encoding the artificial biotin acceptor peptide to the N-terminus of the obelin cDNA gene. The application of the fusion protein as a label in immunoassay was demonstrated. ... 

Tatsumi H, Fukuda S, Kikuchi M, Koyama Y. Construction of biotinylated firefly luciferase using biotin acceptor peptides. Anal Biochem 1996 243 176-80. 

Biotin acceptor peptides >15 Biotin Biotin ligase, ATP Covalent and irreversible Intracellular, cell surface Allows use of derivatized streptavidins to label cell surface proteins [31]... 

Biotin acceptor peptides >15 Keto isostere of biotin Biotin ligase, ATP, hydrazide derivatives Covalent and reversible Cell surface Two-step labeling required [32]... 

Throughout this review, terms such as unfolded, loosely folded, or export-competent state are bandied about (no doubt distressing some disciples of protein structure). These terms are not meant to imply a single definable conformation, or the absence of a specific secondary or tertiary structure. In fact, several lines of evidence indicate that a translocation-competent precursor can contain considerable structure. For example, efficient signal peptide function requires that the hydrophobic core assume an a-helical conformation (Jones et ai, 1990), and evidence has been presented that enzymatically active DHFR (Freudl et al., 1988) and a fully folded biotin acceptor domain (Reed and Cronan, 1991)—or conformers of these proteins that are in equilibrium with the native states—can be translocated across the cytoplasmic membrane of E. coli in vivo. 

Another general approach for the labeling of fusion proteins, which conceptually resembles the labeling of CP-fusion proteins is the biotinylation of so-called acceptor peptides by biotin ligase [31, 58]. The biotinylation of fusion proteins by itself is a valuable modification, as numerous streptavidin-and avidin-based probes and materials are commercially available. However,... 

The peptide group can be fixed at the ice lattice with p = 0127. Ice-like distances of the H-bond acceptors are in different molecules for example Biotin. Thymine, Triglyceride, 1,4-Quinone etc.141. Hechter gave a model for biologic membranes too142. In the membrane surface Hechter assumes a water double hydrate lamella of 4.9 A thickness in which K+ could be placed, but Na+ would be too big. 

Kinase or phosphatase assays based on the AlphaScreen principle are similar to TR-FRET assays in that they usually require a biotinylated substrate peptide and an anti-phosphoserine or tyrosine antibody. These two reagents are sandwiched between biotin and protein A-functionalized acceptor and donor beads. A kinase assay would show an enzyme-dependent increase in antibody binding (and thus signal) over time and a phosphatase assay would show an enzyme-dependent decrease in antibody binding over time. In some cases, the phosphorylation of an epitope will block the antibody binding and thus a phosphatase assay in principle can be constructed as a signal increase assay (Von Leoprichting and Kumpf, 2004 Warner et al., 2004). 

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