Search results for "Isoelectric Point"
showing 10 items of 65 documents
Protein transport through gold-coated, charged nanopores: Effects of applied voltage
2006
The flux of bovine serum albumin and bovine hemoglobin through charged nanopores inside polymeric membranes is analysed as a function of the applied voltage to the nanopore surface, the solution ionic strength and pH. The electrostatic interaction of the protein with the nanopore surface gives low transport rates except at the protein isoelectric point and the minimum of the effective, voltage-induced nanopore charge. This electrostatic sieving effect allows for the separation of proteins with similar molecular weights.
Covalent modificaition of juvenile hormone binding proteins by photoaffinity labeling: An unexpected gel shift effect
1994
The 32 kD juvenile hormone binding protein (JHBP) and two 80 kD proteins in larval Manduca sexta hemolymph were labeled with [3H]FDK, a photoaffinity analog of methyl farnesoate (MF). The labeling could be completely displaced by a 30-fold excess of either MF or JH II, demonstrating that [3H]FDK binds specifically to the JH binding sites of the 32 kD JHBP and the 80 kD proteins. In addition, a high molecular-mass protein was labeled with [3H]FDK; labeling could be displaced by excess MF but not by JH II, demonstrating the selectivity in binding MF. The 32 kD JHBP also appeared to weakly bind the potent juvenoid, methoprene, at the JH binding site. Covalent modification by [3H]FDK induced a …
Solution Properties and Potential Biological Applications of Zwitterionic Poly(ε-N-methacryloyl-l-lysine)
2013
Poly(e-N-methacryloyl-l-lysine) (PMALys) was synthesized by free radical polymerization yielding a zwitterionic polymer with Mw = 721 000 g mol–1. The polymer dissolves in pure water as well as in aqueous salt solution up to 5 M NaClO4 and over wide range of pH values (1.3 ≤ pH ≤ 12.7) as single chains without any sign for aggregate formation. The zwitterionic polymer shows an expanded random coil structure at and close to isoelectric conditions and further expands upon addition of acid and base, respectively. The polymer fulfills four major prerequisites for a promising nano carrier in potential biomedical applications: (1) It is biocompatible, indicated by a low cytotoxicity. (2) It does …
The second component of human complement: Detection of two hemolytic forms in plasma by pH Variation
1988
The second component of human complement (C2) in pseudoglobulin prepared from normal plasma eluted as a single peak at high conductivity (30 mS) and pH 4.5 from the cationic exchangers S-Sepharose or Mono S in the Fast Protein Liquid Chromatography (FPLC) System. The C2 was stable at pH 4.5 and 0 degrees C if enzyme inhibitors were used and the pH was raised to 6.0 after elution from the columns. After rechromatography on Mono S in the FPLC System at the median isoelectric point of 5.5 or pH 6.0, the C2 eluted as two distinct hemolytic forms: the first peaked at 16 mS, the second at 30 mS. The two forms of C2 did not correlate with the allotypic variant of C2 in individual, normal human pla…
Application of CEC procedures for the analysis of synthetic peptides: characterization of linear immunogenic peptides that mimic a HIV-1 gp120 epitope
2002
In this study, we describe the application of a new analytical procedure based on capillary electrochromatographic(CEC) techniques for the characterization of different basic and acidic peptides using isocratic eluent conditions containing acetonitrile and ammonium acetate buffers of different molarities between pH 3.8 and 5.2. In particular,10 immunogenic peptide analogs with isoelectric points ranging from 3.7 to 10.1 were investigated; nine of these peptides, 1-9, were truncated analogs of the parent peptide, 10, which is a peptidomimetic related to a HIV-1 gp120 epitope. Several of these peptides have the propensity to form alpha-helical secondary structures in solution. Electrochromato…
Methods for Separating Native Enzymes
1994
In the course of electrophoresis the stability of an enzyme depends on such conditions as (a) pH-value, (b) ion strength and ion species, (c) effector molecules, (d) temperature and (e) properties of the separation matrix. These parameters were empirically optimized for starch gel electrophoresis [1–3] and cellulose acetate electrophoresis [4, 5] when analyzing predominantly animal and human specimen. A major advantage of these types of separation media is that practically every buffer system can be used to separate enzymes whereas in disc-gel electrophoresis [6–8] the number of applicable buffer systems is limited. When using isoelectric focusing to separate native enzymes no buffer choice…
Contribution of Protein Flexibility to the Foaming Properties of Casein
1990
The effect of biopolymer flexibility on the foaming properties of casein was investigated. Flexibility was altered by: (1) chemical modification (covalent binding of a monosaccharide on the lysyl residues) or (2) pH change. Electron Spin Resonance was used to measure the reorientational frequency of casein residues labeled with nitroxide radicals. High levels of glycosylation induced increased protein flexibility and improved the foaming capacity. Good agreement was observed between higher values of flexibility and improved surface properties near the isoelectric point.
Nucleoside phosphotransferase of chick embryo
1979
This paper describes a purification procedure and some properties of a nonspecific nucleoside phosphotransferase of chick embryo, an activity which catalyzes the transfer of chick embryo, an activity which catalyzes the transfer of the phosphate ester from a deoxyribonucleotide or a pyrimidine ribonucleotide to a deoxyribonucleoside acceptor. The enzyme is very unstable to heat, dilution and dialysis and it is almost entirely inactivated by DEAE-cellulose chromatography or gel filtration. A marked enhancement in its stability is caused by numerous nucleotides. In these experiments at least 920-fold purification was obtained by using dTTP (50 microM) as nucleotide protector. The enzyme, puri…
Biochemical identification and tissue-specific expression patterns of keratins in the zebrafish Danio rerio
1998
We have identified a number of type I and type II keratins in the zebrafish Danio rerio by two-dimensional polyacrylamide gel electrophoresis, complementary keratin blot-binding assay and immunoblotting. These keratins range from 56 kDa to 46 kDa in molecular mass and from pH 6.6 to pH 5.2 in isoelectric point. Type II zebrafish keratins exhibit significantly higher molecular masses (56-52 kDa) compared with the type I keratins (50-48 kDa), but the isoelectric points show no significant difference between the two keratin subclasses (type II: pH 6.0-5.5; type I: pH 6.1-5.2). According to their occurrence in various zebrafish tissues, the identified keratins can be classified into "E" (epider…
The Anaphylatoxic Peptide C3a of Guinea Pig Complement
1978
Abstract Highly purified guinea pig C3a was obtained after specific cleavage of isolated C3 by the alternative pathway enzyme VF-B in a one step procedure. It turned out to be a low molecular weight peptide with basic character (M.W. 9500; isoelectric point above 9.4). C3a represents an antigenetic determinant of its own in the native C3 molecule, different from the B determinant. Guinea pig C3a is resistant to 100°C for 10 minutes. Its smooth muscle contracting activity can be destroyed by trypsin and carboxypeptidase B. These findings indicate that guinea pig C3a is quite similar to human C3a.