ANTIGENICITY OF 5-HYDROXYINDOLE-3-ACETIC ACID, A DERIVATIVE OF SEROTONIN: I. PREPARATION OF PROTEIN CONJUGATES OF 5-HYDROXYINDOLE-3-ACETIC ACID

1967 ◽  
Vol 45 (11) ◽  
pp. 1681-1688 ◽  
Author(s):  
N. S. Ranadive ◽  
A. H. Sehon
Keyword(s):  
Acetic Acid ◽  
Aminocaproic Acid ◽  
Acid Anhydride ◽  
Rabbit Serum ◽  
Amino Groups ◽  
Serum Albumins ◽  
Peptide Bonds ◽  
Protein Conjugates ◽  
Γ Globulins

5-Hydroxyindole-3-acetic acid (5HIAA) was coupled to human, bovine, and rabbit serum albumins, to bovine γ-globulins, and to ε-aminocaproic acid (ACA) through the formation of peptide bonds. For this purpose N,N′-dicyclohexylcarbodiimide was used to prepare the intermediate acid anhydride of 5HIAA, which was subsequently reacted with the amino groups of proteins or ACA. Evidence is presented for the coupling of (i) about 18 residues of 5HIAA per molecule of albumin, (ii) about 30 residues per globulin molecule, and (iii) 1 residue per molecule of ACA.

scholarly journals FREE AMINO GROUPS OF EQUINE γ-GLOBULINS AND A SPECIFIC ANTIBODY

1955 ◽  
Vol 216 (2) ◽  
pp. 621-624
Author(s):  
Mary L. McFadden ◽  
Emil L. Smith
Keyword(s):  
Specific Antibody ◽  
Free Amino ◽  
Amino Groups ◽  
Γ Globulins

Diagonal Chromatography for the Selective Purification of Tyrosyl Peptides

1974 ◽  
Vol 52 (11) ◽  
pp. 1013-1017 ◽  
Author(s):  
William H. Cruickshank ◽  
Barry L. Malchy ◽  
Harvey Kaplan
Keyword(s):  
Acetic Acid ◽  
Stationary Phase ◽  
Enzymatic Digestion ◽  
Acid Water ◽  
Chromatographic Purification ◽  
Amino Groups ◽  
Chromatographic System ◽  
Original Direction ◽  
Acetic Acid Water ◽  
Citraconic Anhydride

Thiolysis of an O-dinitrophenyl-tyrosyl peptide results in an increased solubility in the stationary phase of a n-butanol – acetic acid – water – pyridine (15:3:12:10) (BAWP) paper chromatographic system. It is shown that this property can be used to form the basis of a diagonal paper chromatographic purification of tyrosyl peptides from enzymatic digests of proteins. The amino groups of the protein are first reacted with citraconic anhydride and then the citraconyl protein is reacted with 1-fluoro-2,4-dinitrobenzene. The dinitrophenyl-citraconyl protein is subjected to enzymatic digestion, applied to a strip of Whatman 3 MM paper, thiolyzed with 5% 2-mercaptoethanol in acetone, and subjected to chromatography using BAWP as solvent. A guide strip is removed, thiolyzed with 5% 2-mercaptoethanol in 25% pyridine, and resubjected to chromatography in BAWP at right angles to the original direction of chromatography. The tyrosyl peptides are displaced off the diagonal towards the origin. The off-diagonal peptides are isolated from the original chromatogram by thiolysis and chromatography using the diagonal chromatogram to locate the positions of the dinitrophenyl-tyrosyl peptides.

scholarly journals Structural and immunologic characterization of bovine, horse, and rabbit serum albumins

2012 ◽  
Vol 52 (3-4) ◽  
pp. 174-182 ◽  
Author(s):  
Karolina A. Majorek ◽  
Przemyslaw J. Porebski ◽  
Arjun Dayal ◽  
Matthew D. Zimmerman ◽  
Kamila Jablonska ◽  
...  
Keyword(s):  
Rabbit Serum ◽  
Serum Albumins

scholarly journals Protein thiolation and reversible protein-protein conjugation. N-Succinimidyl 3-(2-pyridyldithio)propionate, a new heterobifunctional reagent

1978 ◽  
Vol 173 (3) ◽  
pp. 723-737 ◽  
Author(s):  
J Carlsson ◽  
H Drevin ◽  
R Axén
Keyword(s):  
Gamma Globulin ◽  
De Novo ◽  
Ester Group ◽  
Alpha Amylase ◽  
Amino Groups ◽  
Protein Conjugates ◽  
Disulphide Bonds ◽  
Concomitant Reduction ◽  
Antibody Conjugate ◽  
Aliphatic Thiols

A heterobifunctional reagent, N-succinimidyl 3-(2-pyridyldithio)propionate, was synthesized. Its N-hydroxysuccinimide ester group reacts with amino groups and the 2-pyridyl disulphide structure reacts with aliphatic thiols. A new thiolation procedure for proteins is based on this reagent. The procedure involves two steps. First, 2-pyridyl disulphide structures are introduced into the protein by the reaction of some of its amino groups with the N-hydroxysuccinimide ester sie of the reagent. The protein-bound 2-pyridyl disulphide structures are then reduced with dithiothreitol. This reaction can be carried out without concomitant reduction of native disulphide bonds. The technique has been used for the introduction of thiol groups de novo into ribonuclease, gamma-globulin, alpha-amylase and horseradish peroxidase. N-Succinimidyl 3-(2-pyridyldithio)propionate can also be used for the preparation of protein-protein conjugates. This application is based on the fact that protein-2-pyridyl disulphide derivatives (formed from the reaction of non-thiol proteins with the reagent) react with thiol-containing proteins (with native thiols or thiolated by, for example, the method described above) via thiol-disulphide exchange to form disulphide-linked protein-protein conjugates. This conjugation technique has been used for the preparation of an alpha-amylase-urease, a ribonuclease-albumin and a peroxidase-rabbit anti-(human transferrin) antibody conjugate. The disulphide bridges between the protein molecules can easily be split by reduction or by thiol-disulphide exchange. Thus conjugation is reversible. This has been demonstrated by scission of the ribonuclease-albumin and the alpha-amylase-urease conjugate into their components with dithiothreitol. N-Succinimidyl 3-(2-pyridyldithio)propionate has been prepared in crystalline form, in which state (if protected against humidity) it is stable on storage at room temperature (23 degrees C).

Abolition by Myosin and Heavy Meromyosin of the Inhibitory Effect of Smooth-Muscle Actomyosin Antibodies on Cell Aggregation In Vitro

1973 ◽  
Vol 12 (2) ◽  
pp. 631-639
Author(s):  
R. B. KEMP ◽  
B. M. JONES ◽  
U. GRÖSCHEL-STEWART
Keyword(s):  
Smooth Muscle ◽  
Striated Muscle ◽  
Cell Aggregation ◽  
Inhibitory Effect ◽  
Regulatory Function ◽  
Rabbit Serum ◽  
Heavy Meromyosin ◽  
Γ Globulins ◽  
Dissociated Cells

The ability of anti-chicken smooth-muscle actomyosin γ-globulins (anti-GAM) to inhibit the aggregation of dissociated cells from the skeletal muscle and liver of chick embryos was abolished by pretreatment of the anti-GAM with either myosin or heavy meromyosin (HMM). When the same cells were treated with HMM at a concentration of 1 mg per 2 x 106 cells/ml Eagle's MEM they aggregated as readily as untreated cells. The negative electrophoretic mobility of the embryonic chick fibroblastic cells was significantly reduced by the globulin fraction of anti-GAM but not of HMM-treated anti-GAM or non-immunized rabbit serum. Anti-chicken striated muscle actomyosin γ-globulins slightly reduced negative mobility but HMM had no effect. The experiments show that the inhibitory effect on cell aggregation of anti-GAM preparations is produced by the anti-myosin antibodies. They also provide support for the theory that a surface-localized myosin-like protein has a regulatory function in cell adhesion.

Sign in / Sign up

Export Citation Format

Share Document