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Biomedical subjects

J C Jaton

Publications and source records attributed to J C Jaton.

At least 55 records · Page 3Linked to original sources
PubMed1982

Biosynthesis and structure of membrane and secretory immunoglobulins.

Almost all of the body's extracellular immunoglobulin (Ig) is derived from Ig-secreting plasma cells of lymphoid tissues. The secreted material is a heterogeneous mixture of different classes and specificities. Lymphoid tissues also contain a large number of essentially non-secretory cells--B lymphocytes--which bear Ig firmly associated with their plasma membranes. Ig molecules thus exist in two functionally different forms, as membrane-bound antigen receptors on the surface of B lymphocytes on the one hand, and as humoral secreted Ig antibodies on the other. On B cells, membrane-bound heavy chains have an apparent mol. wt. slightly larger than that of secreted heavy chains from plasma cells. Membrane-bound but not secreted heavy chains bind detergents, thus suggesting the presence of a hydrophobic region in membrane-bound heavy chains, which is absent in secreted heavy chains. Most investigations have dealt with immunoglobulin M. The two types of IgM heavy chains differ at their carboxy termini. Recent investigations at the nucleic acid level demonstrate that membrane-associated mu chains contain a 41-residue hydrophobic tail adjacent to the last constant domain, whereas secretory mu chains contain a 20-residue hydrophilic tail. At the present time, evidence is accumulating that all membrane-bound Ig heavy chain classes may contain similar hydrophobic structures necessary for anchorage of the molecules into the lipid bilayer.

Animals↗
PubMed1982

Allotypes in Basilea rabbits.

Basilea rabbits produce immunoglobulin molecules, practically all of which have lambda light chains rather than kappa chains. This variant strain was derived form a homozygous (b9/b9) male. Sensitive serological methods revealed that at least some homozygous bas/bas individuals possess traces (about 100 ng/ml) of b9 molecules. This level usually increases to almost 1 microgram/ml upon hyperimmunization with pneumococcal or streptococcal vaccines. One exceptional rabbit, with 50 micrograms/ml of b9 molecules, was found. In spite of the presence of b9 molecules in early pre-immune bleeds, homozygous bas/bas rabbits produce strong anti-b9 antibodies; i.e., they are capable of making autoantibodies. These anti-b9 allotypic antisera were not distinguishable by our methods from routinely produced anti-b9.

Animals↗
PubMed1982

Structural differences between heavy chains of secreted and membrane-bound IgM of a human lymphoblastoid cell line.

The human lymphoblastoid cell line BL was shown to synthesise three distinct molecular species of immunoglobulin M heavy chains: membrane-bound (micrometer). intracellular (micro i) and secreted (microseconds) micro-chains. Only the membrane-bound form could be labeled with a lipophilic photoactivatable nitrene reagent. Analysis of their constituent CNBr fragments and carboxypeptidase A and B digestions of their C-terminal tails suggest that the CNBr peptide pattern of microseconds and micrometer, though similar, is not identical, and that amino acids released at the C-termini of the chains are different. The data confirm recent observations in human and murine systems be showing that the membranes-associated human micro-chain contains a hydrophobic segment, consistent with its anchorage into the lipid bilayer of the plasma membrane and a C-terminal amino acid sequence different from that of the secretory micro-chain.

Amino Acid Sequence↗
PubMed1982

BASILEA rabbits express two types of immunoglobulin light chains: lambda and kappa-like.

In the variant rabbit strain BASILEA, immunoglobulin G were shown to contain two distinct populations of IgG molecules whose light (L) chains belonged to the known lambda isotype and to a new kappa-like type. These two L chains differed from each other by electrophoretic, chemical, and antigenic properties. The kappa-like L chain fraction showed (i) an acid-labile Asp-Pro bond at the end of the joining region and (ii) a tryptic peptide, whose amino acid sequence of the NH2-terminal 15 residues was identical to the homologous constant (C) region sequence of b9 kappa chain with the exception of the residue in position 70, which is asparagine in the kappa-like chain instead of the characteristic half-cystine residue in all L chains of kappa B type expressing b4, b5, b6, or b9 allotypes. The data suggest that the kappa-like L chain component does not contain the C region half-cystine residue involved in the formation of the extra variable (V) region-C region disulfide bridge in L chains of the kappa B [Rejnek, J., Appella, E., Mage, R. G. & Reisfeld, R. A. (1969) Biochemistry 8, 2712-2718]. The partial NH2-terminal amino acid sequence of the C region of the kappa-like L chain was shown to be markedly different from b4, b6, or b9 region sequences and from rabbit lambda C region sequence. Taken together, the chemical data suggest that the kappa bas component represents a new subtype of kappa chain. A rabbit alloantiserum made against bas IgG and adsorbed with IgG fractions showing b4, b5, b6, b95, and b96 L chain allotypes appeared to be directed against the kappa-like L chain component of BASILEA IgG exclusively. All BASILEA animals expressed IgG molecules containing kappa-like chains; in contrast, IgG molecules derived from the standard domestic rabbit did not react with this antiserum.

Amino Acid Sequence↗
PubMed1981

The primary structure of the constant region of Basilea-rabbit immunoglobulin lambda-chains.

The amino acid sequence of the constant region of the immunoglobulin lambda-chain of the rabbit (C lambda) has been determined. This chain was obtained from a homogeneous anti-(type II pneumococcal polysaccharide) antibody fraction raised in Basilea rabbit 4548. The C lambda-region sequence was established by the analysis of tryptic and some overlapping chymotryptic peptides and partly by homology with known C lambda-region sequences of lambda-chains from other species. Peptides were isolated by a combination of methods including high-voltage paper electrophoresis, gel filtration and high-pressure liquid chromatography. The peptides were subjected to automated Edman degradation in the presence of Polybrene. The sequence showed no evidence of heterogeneity. Large interspecies similarities, as well as amino acid interchanges, are apparent among various C lambda regions; the degrees of homology between rabbit C lambda region and the C lambda region from human, porcine and murine chains are 72.6, 71 and 72% respectively. The similarity of lambda-chains of different species is much greater than that found between rabbit kappa- and lambda-chains (about 34% homology). Genetic implications of these results are discussed.

Amino Acid Sequence↗
PubMed1980

Biosynthesis of two forms of IgM heavy chains by normal mouse B lymphocytes. Membrane and secretory IgM.

A study of the biosynthesis of IgM by purified mouse spleen lymphocytes showed that these cells synthesize both 8 S membrane IgM and 19S secretory IgM, which is identical with plasma cell IgM except in its kinetics of processing, assembly, and secretion. The heavy (mu) chains of these two types of lymphocyte IgM differ in their ultimate fate, in processing, isoelectric point, and peptide composition. The separate precursors of the two mu chains have very similar mobilities in sodium dodecyl sulfate polyacrylamide gel electrophoresis, but they can be distinguished by the use of endoglucosaminidase H (endo-H) to remove core sugars, by two-dimensional electrophoresis, and by one-dimensional gel analysis in pulse-chase experiments. CNBr peptide patterns of intracellular "secretory" mu chains of lymphocytes and plasma cells were similar, but membrane mu chains had a COOH-terminal peptide different in structure from that of secreted mu chains, with a higher apparent molecular weight.

Animals↗
PubMed1980

Preparation and characterization of chemically defined oligomers of rabbit immunoglobulin G molecules for the complement binding studies.

Pure dimers, trimers, tetramers and pentamers of rabbit non-immune IgG (immunoglobulin G) or antibody IgG were prepared by polymerization in the presence of the bifunctional cross-linking reagent dithiobis (succinimidylpropionate). Oligomerization was performed either in the presence of polysaccharide antigen and specific monomeric antibody (method A) or by random cross-linking of non-immune rabbit IgG in the absence of antigen (method B). By repeated gel-filtration chromatography, samples prepared by both methods exhibited a single band in analytical sodium dodecyl sulphate/polyacrylamide-gel electrophoresis. The electrophoretic mobilities of samples prepared by method A were slightly greater than those for the corresponding samples prepared by method B. This might suggest a role played by antigen in the orientation of IgG molecules within the clusters, which may be more compact than those formed by random cross-linking. The average numbers of cross-linker molecules per oligomer varied between 3 and 6 for clusters made by method A and between 1 and 3 for clusters made by method B. Ultracentrifugal analyses of the oligomers yielded sedimentation coefficients (S20,w) of 9.6S for the dimer, 11.2S for the trimer, 13.6S for the tetramer and 16.1S for the pentamer. Comparison of the observed sedimentation coefficients with those predicted by various hydrodynamic models suggested these oligomers possessed open and linear structures. Reduction of the cross-linking molecules converted oligomers into monomeric species of IgG. C.d. spectra of some oligomers studied in the range 200-250 nm were essentially the same as that of monomeric IgG molecules, thus strongly suggesting no major conformation changes in IgG molecules within clusters. These oligomers were found to be stable for up to 2 months when stored at -70 degrees C.

Animals↗
PubMed1980

Dimeric, trimeric and tetrameric complexes of immunoglobulin G fix complement.

The binding of pure dimers, trimers and tetramers of randomly cross-linked non-immune rabbit immunoglobulin G to the first component and subcomponent of the complement system, C1 and C1q respectively, was studied. These oligomers possessed open linear structures. All three oligomers fixed complement with decreasing affinity in the order: tetramer, trimer, dimer. Complement fixation by dimeric immunoglobulin exhibited the strongest concentration-dependence. No clear distinction between a non-co-operative and a co-operative binding mechanism could be achieved, although the steepness of the complement-fixation curves for dimers and trimers was better reflected by the co-operative mechanism. Intrinsic binding constants were about 10(6)M-1 for dimers, 10(7)M-1 for trimers and 3 X 10(9)M-1 for tetramers, assuming non-co-operative binding. The data are consistent with a maximum valency of complement component C1 for immunoglobulin G protomers in the range 6-18. The binding of dimers to purified complement subcomponent C1q was demonstrated by sedimentation-velocity ultracentrifugation. Mild reduction of the complexes by dithioerythritol caused the immunoglobulin to revert to the monomeric state (S20,w = 6.2-6.5S) with concomitant loss of complement-fixing ability.

Animals↗
PubMed1980

Antigen-independent binding of IgG dimers to C 1 q as studied by sedimentation equilibrium, complement fixation and electron microscopy.

Rabbit anti-SII pneumococcal polysaccharide IgG antibody was cross-linked by dithiobis (succinimidyl propionate). The IgG dimers were visualized by electronmicroscopy and resembled two Y-shaped structures connected in random orientations. The three arms of the Y's were of equal length. The six globular domains of the IgG protomers could be seen, but the Fab arms and Fc stems could not be distinguished from each other. Electron microscopy revealed that the dimers were bound to the globular heads of C 1 q. From the dependence of thee weight average molecular weight on dimer concentration, and equilibrium constant of about 10(6) M(-1) was derived for the binding of dimers to C 1 q. The number of IgG dimers which could be accommodated at a single C 1 q molecule was derived to be three. The data do not allow a clear distinction between noncooperative and cooperative binding. The binding equilibrium was independent of whether the IgG dimers were liganded with an SII nonasaccharide hapten or not. The results are at variance with an allosteric mechanism of the action of antigen in C1 activation. They lend support to the association hypothesis of complement activation. The data suggest that clusters of about six IgG molecules, connected by a multivalent antigen or arranged at the cell surface, are recognized by C 1 q with a binding constant of about 10(10) M(-1).

Animals↗
PubMed1980

Antibody diversity in amphibians: evidence for the inheritance of idiotypic specificities in isogenic Xenopus.

Two families of isogenic Xenopus and the progeny from one individual of each family were immunized with dinitrophenylated keyhole limpet hemocyanin. Antisera were raised in mice and in guinea pigs against purified antibodies from one individual of the first generation of each family. These were rendered anti-idiotypic by absorption with immunoglobulins of unimmunized isogenic frogs. In the first generation, antibodies from individuals of each family exhibited a high degree of idiotypic cross-reactivity. Antibodies from individuals of the second generation displayed idiotypic cross-reactivity, but to a lesser extent. Some idiotypic cross-reactivity was seen between members of one family with members of the other family: no sharing of idiotypic specificity was detected among anti-DNP antibodies isolated from individual outbred frogs. Isoelectric focusing (IEF) patterns of the anti-DNP antisera showed a high degree of restriction and overlap within members of each family and across two generations. Furthermore, the N-terminal sequence of the L chains from X. laevis anti-DNP antibodies was unique, and there was a low degree of structural heterogeneity in the H chain N-terminal region. A correlation between the IEF banding pattern and idiotypic reactivity was apparent. The data suggest the inheritance of variable region genes through two generations of isogenic Xenopus.

Amino Acid Sequence↗