Generation and expression of a minimal hybrid Ig-receptor formed between single domains from proteins L and G

Harrison, S.; Housden, Nicholas G.; Bottomley, Stephen P.; Cossins, Aimee J.; Gore, Michael G.

doi:10.1016/j.pep.2007.11.007

Cited by 5 publications

(3 citation statements)

References 24 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The immunodiffusion assay was performed as previously reported ( 37 ) using 0.5% agarose gel that was melted in potassium phosphate buffer, pH 6.65, containing 1% (w/v) β-OG. Approximately 4 ml of gel was poured into a 35-mm Petri dish.…”

Section: Methodsmentioning

confidence: 99%

“…To investigate this we carried out ITC experiments with different regions of ColN. To further investigate the OBS of ColN we divided the putative sequence into two peptides: ColN [2][3][4][5][6][7][8][9][10][11][12][13][14][15][16][17][18][19] and ColN [20][21][22][23][24][25][26][27][28][29][30][31][32][33][34][35][36][37][38] , of which only ColN [2][3][4][5][6][7][8][9][10][11][12][13][14][15]…”

Section: Coln Has Distinct Ompf-binding Sitesmentioning

confidence: 99%

See 1 more Smart Citation

Bifurcated binding of the OmpF receptor underpins import of the bacteriocin colicin N into Escherichia coli

Jansen

Inns

Housden

et al. 2020

Journal of Biological Chemistry

View full text Add to dashboard Cite

Colicins are Escherichia coli–specific bacteriocins that translocate across the outer bacterial membrane by a poorly understood mechanism. Group A colicins typically parasitize the proton-motive force–linked Tol system in the inner membrane via porins after first binding an outer membrane protein receptor. Recent studies have suggested that the pore-forming group A colicin N (ColN) instead uses lipopolysaccharide as a receptor. Contrary to this prevailing view, using diffusion-precipitation assays, native state MS, isothermal titration calorimetry, single-channel conductance measurements in planar lipid bilayers, and in vivo fluorescence imaging, we demonstrate here that ColN uses OmpF both as its receptor and translocator. This dual function is achieved by ColN having multiple distinct OmpF-binding sites, one located within its central globular domain and another within its disordered N terminus. We observed that the ColN globular domain associates with the extracellular surface of OmpF and that lipopolysaccharide (LPS) enhances this binding. Approximately 90 amino acids of ColN then translocate through the porin, enabling the ColN N terminus to localize within the lumen of an OmpF subunit from the periplasmic side of the membrane, a binding mode reminiscent of that observed for the nuclease colicin E9. We conclude that bifurcated engagement of porins is intrinsic to the import mechanism of group A colicins.

show abstract

Section: Methodsmentioning

confidence: 99%

Section: Coln Has Distinct Ompf-binding Sitesmentioning

confidence: 99%

Bifurcated binding of the OmpF receptor underpins import of the bacteriocin colicin N into Escherichia coli

Jansen

Inns

Housden

et al. 2020

Journal of Biological Chemistry

View full text Add to dashboard Cite

show abstract

“…In 2008, we identified more than 500 articles after searching Web of Science, PubMed, SciDir and OVID databases using ‘isothermal AND titration AND calorimetry’ or ITC or ‘Isothermal Titration Calorimetry’ search terms. Following the format of previous annual surveys these have been classified into the following categories: References cited in the text and review articles 1–82. Protein–protein and protein–peptide interactions 83–222. Protein–small ligand or protein–drug interactions 223–321. Protein/peptide metal interactions 322–355. Protein/peptide nucleic acid interactions 356–387. Protein/peptide lipid interactions …”

Section: Introductionmentioning

confidence: 99%

Survey of the year 2008: applications of isothermal titration calorimetry

Falconer

Penkova

Jelesarov

et al. 2010

J of Molecular Recognition

View full text Add to dashboard Cite

Isothermal titration calorimetry (ITC) is a fast, accurate and label-free method for measuring the thermodynamics and binding affinities of molecular associations in solution. Because the method will measure any reaction that results in a heat change, it is applicable to many different fields of research from biomolecular science, to drug design and materials engineering, and can be used to measure binding events between essentially any type of biological or chemical ligand. ITC is the only method that can directly measure binding energetics including Gibbs free energy, enthalpy, entropy and heat capacity changes. Not only binding thermodynamics but also catalytic reactions, conformational rearrangements, changes in protonation and molecular dissociations can be readily quantified by performing only a small number of ITC experiments. In this review, we highlight some of the particularly interesting reports from 2008 employing ITC, with a particular focus on protein interactions with other proteins, nucleic acids, lipids and drugs. As is tradition in these reviews we have not attempted a comprehensive analysis of all 500 papers using ITC, but emphasize those reports that particularly captured our interest and that included more thorough discussions we consider exemplify the power of the technique and might serve to inspire other users.

show abstract

Analysis and Purification of Antibody Fragments Using Protein A, Protein G, and Protein L

Griep¹,

McDougall²

2010

Antibody Engineering

View full text Add to dashboard Cite

Generation and expression of a minimal hybrid Ig-receptor formed between single domains from proteins L and G

Cited by 5 publications

References 24 publications

Bifurcated binding of the OmpF receptor underpins import of the bacteriocin colicin N into Escherichia coli

Bifurcated binding of the OmpF receptor underpins import of the bacteriocin colicin N into Escherichia coli

Survey of the year 2008: applications of isothermal titration calorimetry

Analysis and Purification of Antibody Fragments Using Protein A, Protein G, and Protein L

Contact Info

Product

Resources

About