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Friday, July 17, 2020 | History

2 edition of Peptide-derived sensors with tuned affinity for heparin. found in the catalog.

Peptide-derived sensors with tuned affinity for heparin.

Jimena Celia Sauceda

Peptide-derived sensors with tuned affinity for heparin.

by Jimena Celia Sauceda

  • 40 Want to read
  • 18 Currently reading

Published .
Written in English


About the Edition

A new fluorescent Fmoc-SPPS compatible amino acid that incorporates the halide-sensitive 6-methoxyquinolinium group was synthesized and incorporated in two peptides with known affinity for heparin. Sensor number one, AGf73, showed a 320% increase in fluorescence intensity upon heparin association, with an apparent dissociation constant of 2.6 muM at 100 mM NaCl. AGf73 selectivity was illustrated by the absence of changes in fluorescence intensity in the presence of chondroitin sulfate A. Furthermore, the proposed chloride-dependent sensing mechanism was supported by the fact that fluoride ions did not elicit a significant fluorescence change in the presence or in the absence of heparin. Sensor number two, MK2f-69d-61, was based on the second domain of human midkine, under the same working principle as AGf73, adding an internal fluorescent moiety that would render it ratiometric. Although adequate folding of the second sensor was not achieved, the ratiometric nature of the probe was corroborated experimentally.11This work has been submitted to ChemBioChem and is being revised for publication.

The Physical Object
Paginationxi, 47 leaves :
Number of Pages47
ID Numbers
Open LibraryOL21549708M
ISBN 109780494210437

The sensor was dipped in an aqueous solution of M LiCl, which was stirred with a magnetic stirrer ( Hz), and eventually spiked with heparin ( U mL-1).   For heparin-based purification of EV using magnetic beads,. biotinylated heparin (Sigma-Aldrich, St. Louis, MO) was allowed to bind to MyOne .

Heparin affin regulatory peptide (HARP) is an heparin-binding molecule involved in the regulation of cell proliferation and differentiation. Here, we report that HARP inhibited the biological. Heparin-affinity. μg HSC exosomes were added to μl TSKgel heparin-5PW affinity beads (Tosoh BioScience LLC, King of Prussia, PA) in μl PBS and mixed at 37 o C for 1 hr. The beads were washed 3 times with 1 ml PBS and then split into 10 equal aliquots which were then mixed for 15 mins with μl PBS containing – M NaCl.

In fact, KG appears to encompass only a low-affinity heparin binding site. In contrast, peptides based solely on the high-affinity binding site (KA) displayed much higher affinities for heparin. By CD spectrometry, these high-affinity peptides are chiefly random coil in nature, but low microM concentrations of heparin induce. Affinity chromatography Fig 1. Heparin Sepharose 6 Fast Flow purifies proteins with an affinity for heparin. As a BioProcess resin, the product is available in process-scale quantities. Fig 2. Heparin consists of alternating hexuronic acid (A) and D-glucosamine residues (B). The hexuronic acid can either be D-glucuronic acid (top) or its.


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Peptide-derived sensors with tuned affinity for heparin by Jimena Celia Sauceda Download PDF EPUB FB2

The binding affinity between heparin and VEGF was measured on a heparin sepharose column (HiTrap Heparin HP, 1 ml). The VEGF was injected and eluted under a linear gradient of 0–1 M NaCl in 5 ml sodium phosphate (pH ) at 1 ml/min for 20 min, with UV detection at by:   In this DNA sensor, a peptide derived from the basic region of a bZIP transcription factor, was conjugated with intercalating tripyrrole through an amine linker.

Affinity of the sensor to DNA was enhanced by optimization of the molecular structure design, such as the spacer structure between the peptide and fluorophores [61].Cited by:   The receptor for hyaluronan (HA) 3-mediated motility (RHAMM) features a amino acid HA-binding domain (HABD) that contains two base-rich BX 7 B motifs and possesses an overall helix–turn–helix structure.

This HABD also has significant affinity for heparin, a polysulfated glycosaminoglycan (GAG).RHAMM mediates cell migration and proliferation, and isoforms can be Cited by: Affinity-based release systems use transient interactions to sustain and control the release of a therapeutic from a polymeric matrix.

The most common affinity-based systems use heparin-based scaffolds to sustain the release of heparin-binding proteins, such as fibroblast growth factor-2 (FGF2) and vascular endothelial growth factor (VEGF). However, novel affinity-based systems based on, for Cited by: Equal response on a mass basis occurs with fragments of heparin's as small as Da.

Complexation of heparin's anionic sites by macromolecules that bind heparin with high affinity (e.g. Van Kerkhof et al. (), Van Kerkhof et al. ()developed a heparin sensor with a protamine affinity ligand, based on an ion-sensitive field effect transistor (ISFET) technique with an indirect ion-step method to probe the heparin concentration.

The range of this sensor is to U/ml with an accuracy of U/ml. The ISFET based heparin sensor with a monolayer of protamine as affinity ligand J.C.

van Kerkhof & P. Bergveld MESA Research Institute, University of Twente, P.O. BoxAE Enschede, The Netherlands Tel: [31] 53 Fax: [31] 53 R.B.M.

Schasfoort. First, although the LAL test is sensitive, affinity sensor technology is likely even more sensitive. Second, many factors may influence the LAL test, such as acetic acid and heparin, whereas the biosensor assay could be strictly controlled. Thus, the data from the biosensor assay may more objectively represent the anti-endotoxin activity of.

The comparison of the affinity and kinetic parameters of full-length proteins and their fragments for heparin was performed on 15 and 12 pairs respectively. 87% of full-length proteins had a higher affinity for heparin than their fragments with the.

Light scattering showing heparin clustering by PACAP38 and not by other peptides. (A) Titration of heparin into PACAP38 () or VIP () monitored by turbidity at nm.

Each peak corresponds to the injection of μL of a μM heparin solution (20 mM phosphate, mM NaCl, pH ) into a 1 mL 50 μM peptide solution (20 mM phosphate, The immune response to biomaterial implants critically regulates functional outcomes such as vascularization, transplant integration/survival, and fibrosis.

To create “immunologically smart” materials, the host-material response may be engineered to optimize the recruitment of pro-regenerative leukocyte subsets which mature into corresponding wound-healing macrophages.

The ion-step measuring method was used to determine absolute heparin concentrations in PBS and blood plasma with a Ta2O5 ISFET on to which a monolayer of protamine had been immobilized.

Heparin is a highly negatively charged polysaccharide, which is used clinically to delay the clotting of blood. Protamine acts as an affinity ligand for heparin. USA Home > Product Directory > Molecular Biology > Proteomics > Protein Chromatography > Affinity Chromatography > Heparin Life Science Home Life Science Products.

We report dual therapeutic effects of a synthetic heparin-binding peptide (HBP) corresponding to residues 15–24 of the heparin binding site in BMP4 in a collagen-induced rheumatic arthritis model (CIA) for the first time.

The cell penetrating capacity of HBP led to improved cartilage recovery and anti-inflammatory effects via down-regulation of the iNOS-IFNγ-IL6 signaling pathway in. A review. Heparin is a vital biomol. in widespread clin.

use as an anti-coagulant. Heparin sensors have potential applications in the bedside detection of heparin levels in human blood during surgery, while high-affinity heparin binders may enable the development of effective heparin reversal agents for use in patients once surgery is complete. @article{osti_, title = {Heparin-binding peptide as a novel affinity tag for purification of recombinant proteins}, author = {Morris, Jacqueline and Jayanthi, Srinivas and Langston, Rebekah and Daily, Anna and Kight, Alicia and McNabb, David S.

and Henry, Ralph and. Abstract. Proteoglycan-binding peptides were designed based on consensus sequences in heparin-binding proteins: XBBXBXand XBBBXXBX, where X and B are hydropathic and basic residues, respectively.

Initial peptide constructs included (AKKARA) n and (ARKKAAKA) n (n = 1–6). Affinity coelectrophoresis revealed that low M r peptides (–) had no affinities for low M r heparin, but. Schrader has recently developed a fluorescent polymeric heparin sensor that can quantify this polysaccharide with unprecedented sensitivity (30 nM).

Coupling of boron-carbohydrate interactions with electrostatic attraction in a multivalent manner is responsible for the high affinity of this receptor for its substrate. The sulfated HA macromers bound a heparin-binding protein (i.e., stromal cell-derived factor 1-α, SDF-1α) with an affinity comparable to heparin and did not alter the gelation behavior or.

Competitive binding studies with low-affinity heparin and a heparin tetrasaccharide suggest that PAA binds antithrombin in both the pentasaccharide- and the extended heparin-binding sites, and these results are corroborated by molecular modeling.

Introduction of a Chemical Constraint in a Short Peptide Derived from Human Acidic Fibroblast Growth Factor Elicits Mitogenic Structural Determinants July Journal of Medicinal Chemistry 46(A tumor-derived growth factor that stimulates the proliferation of capillary endothelial cells has a very strong affinity for heparin.

This heparin affinity makes it possible to purify the growth factor to a single-band preparation in a rapid two-step procedure. The purified growth factor is a cationic polypeptide, has a molecular weight of ab, and stimulates capillary endothelial.Heparin is widely recognized for its potent anticoagulating effects, but has an additional wide range of biological properties due to its high negative charge and heterogeneous molecular structure.

This heterogeneity has been one of the factors in motivating the exploration of functional analogues with a more predictable modification pattern and monosaccharide sequence, that can aid in.