Limited cleavage of disulphide bonds of pig gamma-globulin by S-sulphonation

1964 ◽  
Vol 29 (6) ◽  
pp. 1401-1412 ◽  
Author(s):  
F. Franěk ◽  
J. Zikán
Keyword(s):  
Gamma Globulin ◽  
Disulphide Bonds

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).

scholarly journals The disulphide bonds of human and rabbit γ-globulins

1965 ◽  
Vol 97 (2) ◽  
pp. 569-572 ◽  
Author(s):  
R Cecil ◽  
GT Stevenson
Keyword(s):  
Heavy Metal ◽  
Gamma Globulin ◽  
Sh Groups ◽  
Predominant Species ◽  
Disulphide Bonds ◽  
Γ Globulins ◽  
Presence And Absence ◽  
Metal Reagents

1. The reaction of the disulphide bonds of the predominant species of human and rabbit gamma-globulins (the 7s gamma-globulins) with sulphite was studied in the presence and absence of denaturing agents and heavy-metal reagents. 2. The total number of bonds reacting/mol. of mol.wt. 160000 was approx. 18 for human and 20 for rabbit gamma-globulin. 3. Six S.S bonds/mol. of human and 6.5 S.S bonds/mol. of rabbit gamma-globulin reacted with sulphite alone at pH6. These appeared to include all the interchain S.S bonds. 4. The number of free SH groups was less than 0.2/mol. of human and less than 0.3/mol. of rabbit gamma-globulin.

Electron Microscopy of Mycoplasma Pneumoniae

1971 ◽  
Vol 29 ◽  
pp. 258-259 ◽  
Author(s):  
E. S. Boatman ◽  
G. E. Kenny
Keyword(s):  
Electron Microscopy ◽  
Light Microscopy ◽  
Growth Phase ◽  
Hela Cells ◽  
Sulfonic Acid ◽  
Gamma Globulin ◽  
Horse Serum ◽  
Morphological Aspects ◽  
The Family

Information concerning the morphology and replication of organism of the family Mycoplasmataceae remains, despite over 70 years of study, highly controversial. Due to their small size observations by light microscopy have not been rewarding. Furthermore, not only are these organisms extremely pleomorphic but their morphology also changes according to growth phase. This study deals with the morphological aspects of M. pneumoniae strain 3546 in relation to growth, interaction with HeLa cells and possible mechanisms of replication.The organisms were grown aerobically at 37°C in a soy peptone yeast dialysate medium supplemented with 12% gamma-globulin free horse serum. The medium was buffered at pH 7.3 with TES [N-tris (hyroxymethyl) methyl-2-aminoethane sulfonic acid] at 10mM concentration. The inoculum, an actively growing culture, was filtered through a 0.5 μm polycarbonate “nuclepore” filter to prevent transfer of all but the smallest aggregates. Growth was assessed at specific periods by colony counts and 800 ml samples of organisms were fixed in situ with 2.5% glutaraldehyde for 3 hrs. at 4°C. Washed cells for sectioning were post-fixed in 0.8% OSO4 in veronal-acetate buffer pH 6.1 for 1 hr. at 21°C. HeLa cells were infected with a filtered inoculum of M. pneumoniae and incubated for 9 days in Leighton tubes with coverslips. The cells were then removed and processed for electron microscopy.

Ferritin-Conjugated Antibody for the Demonstration of Isoantigens on the Surface of Murine Cells

1968 ◽  
Vol 26 ◽  
pp. 48-49
Author(s):  
T. Aoki ◽  
J. Izard ◽  
U. Hämmerling ◽  
E. de Harven ◽  
L. J. Old
Keyword(s):  
Cell Surface ◽  
Gamma Globulin ◽  
Leukemia Cells ◽  
Labelling Technique ◽  
Direct Labelling ◽  
Labelling Method

Although a variety of viral and cellular antigens have been demonstrated by ferritin-labeled antibody, this technique has not been used to locate isoantigens on the surface of nucleated cells. The recognition of several systems of isoantigens on the surface of thymocytes, lymphocytes and leukemia cells of the mouse and the ease with which these cells can be obtained in free suspension led us to consider the ferritin-labelling method to determine the amount and location of these isoantigens on the cell surface. Because of the problems involved in the direct labelling of mouse gamma globulin by ferritin, we have chosen an indirect labelling technique (i.e. ferritin-conjugated rabbit anti mouse γG)to detect localization of mouse isoantibody.

Gamma-globulin in the prophylaxis of posttransfusion hepatitis

JAMA ◽  
1966 ◽  
Vol 196 (6) ◽  
pp. 471-474 ◽  
Author(s):  
P. V. Holland
Keyword(s):  
Gamma Globulin ◽  
Posttransfusion Hepatitis

Release, Aggregation and Lysis of Human Platelets by Antilymphocyte Globulin and Antiplatelet Serum

1976 ◽  
Vol 36 (02) ◽  
pp. 411-423 ◽  
Author(s):  
Nicholas Lekas ◽  
J. C Rosenberg
Keyword(s):  
Platelet Aggregation ◽  
Gamma Globulin ◽  
Human Platelets ◽  
Plasma Medium ◽  
Release Reaction ◽  
Clinical Events ◽  
Thrombotic Complications ◽  
Platelet Release ◽  
Free Media

SummaryHuman platelets labeled with 51Cr were used to determine the contribution made by platelet lysis to the platelet release reaction and platelet aggregation induced by rabbit antihuman platelet serum (APS) and equine antihuman thymocyte globulin (ATG). Platelets were tested in both plasma (PRP) and non-plasma containing media. Antibodies directed against platelets, either as APS or ATG, induced significant amounts of platelet release and aggregation, as well as some degree of lysis, in the absence of complement. The presence of complement increased platelet lysis and aggregation, but not the release reaction. Non-immune horse gamma globulin produced different responses depending upon whether platelets were investigated in PRP or non-plasma containing media. Aggregation was seen in the latter but not the former. These differences can be explained by the presence of plasma components which prevent non-specific immune complexes from causing platelet aggregation. Since platelets in vivo are always in a plasma medium, one must be wary of utilizing data from platelet studies in synthetic plasma-free media as the basis of explaining clinical events. These observations demonstrate at least two, and possibly three, different mechanisms whereby ATG could activate platelets causing thrombotic complications and thrombocytopenia, i.e., via 1) specific and, 2) non-specific non-lytic pathways and 3) a lytic pathway.

Inhibition of Fibrinoid Deposition by Heparin

1961 ◽  
Vol 06 (01) ◽  
pp. 157-159 ◽  
Author(s):  
Saul B. Gilson
Keyword(s):  
Intravenous Injection ◽  
Gamma Globulin ◽  
Physiological Significance

ConclusionExperimental glomerulitis in rabbits following intravenous injection of gamma globulin was inhibited by heparinization. The physiological and patho-physiological significance of this observation is considered.

Deficiencies and Inhibitors of Thromboplastic Activity

1966 ◽  
Vol 16 (03/04) ◽  
pp. 574-585
Author(s):  
G. F Grannis ◽  
L. A Kazal
Keyword(s):  
Factor Viii ◽  
Time Course ◽  
Gamma Globulin ◽  
Hemophilia A ◽  
Naturally Occurring ◽  
Thrombin Activity ◽  
Activity Curve ◽  
Lipoprotein Complex ◽  
Thromboplastic Activity

SummaryThe effects of hereditary deficiencies of thromboplastic proteins (hemophilia A and B) on the time course of thrombin appearance and disappearance in plasma (the thrombin activity curve, TAC) were compared with the effects of a naturally occurring gamma-globulin inhibitor of thromboplastic activity and with an anti-thromboplastic activity derived from cephalin (phosphatidylserine-lipoprotein complex).Both inhibitors inhibit reactions involving the protein in which hemophilia A plasma is deficient (factor VIII).

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