Search PubMed⌕ Search

Biomedical subjects

S Rudikoff

Publications and source records attributed to S Rudikoff.

At least 109 records · Page 6Linked to original sources
PubMed1979

Crystal structure of galactan-binding mouse immunoglobulin J539 Fab at 4.5-A resolution.

An electron-density map of the mouse galactan-binding immunoglobulin J539 (IgA2,kappa) Fab has been calculated to a resolution of 4.5 A by the method of heavy atom isomorphous replacement with four derivatives. The map has been interpreted with the aid of a computer program which systematically searched for the best fit between the electron-density map and the known coordinates of individual immunoglobulin domains. The quaternary structure of J539 Fab at this resolution appears similar to that of another mouse immunoglobulin, IgA2,kappa Fab, McPC603. The model coordinates for J539 Fab should allow us to proceed directly to a high-resolution structure determination without further heavy atom isomorphous replacement.

Animals↗
PubMed1979

Structural evidence for a polymorphic or allelic form of the heavy chain variable region.

The heavy (H) chains of anti-phosphocholine (PC) antibodies from C57BL/6J and CBA/J were sequenced through the N-terminal 36 residues and compared with previously published sequences of A/J anti-PC antibody and BALB/c PC-binding myeloma proteins T15, M603, and M511. Each of these antibody preparations contained molecules having light (L) chains and idiotypic determinants of T15, M511, and M603 indicating the presence of at least three different anti-PC antibodies in each pool. The structures of the C57BL/6J and CBA/J H chains each revealed a single sequence from positions 1 to 36 (which includes the first complementarity determining region (CDR), and they were identical. The first CDR was identical to that previously found for BALB/c and A/J indicating that this portion of these antibody molecules is highly conserved throughout inbred mice and is probably critical to PC-binding. A surprising finding was that both C57NL/6 and CBA sequence differed from the BALB/c and A/J sequences at two positions, residue 14 and 16. Since each of these strains differs at the allotype locus, the data indicates that the evolution of allotypy in mice occurred after variable region diversity for the particular genes.

Alleles↗
PubMed1978

k Chain variable regions from three galactan binding myeloma proteins.

A series of seven BALB/c myeloma proteins has been identified with binding specificity for antigens containing beta(1 leads to 6)-D-galactopyranosyl moieties. We have determined the primary amino acid sequence of the first 108 residues from the light chains of three of these proteins. The framework portions of the variable regions of these three light chains are identical with residue 100 at which position three different amino acids are found in the three chains. An additional interchange was found at position 106 in one of the proteins. Based on recent DNA sequence studies suggesting that the variable region ends at residue 97, these substitutions indicate the possible existance of multiple genes coding for the region beginning at residue 98 and continuing toward the carboxy terminus. A single amino acid interchange was observed in complementarity determining regions occurring in L3. This substitution (Ile-Trp) would require changes in all three codon bases to produce the respective amino acids if one were derived from the other. Two of these chains are thus indistinguishable for their first 100 amino acids and are the first pair of k chains to exhibit complete identity over their variable regions.

Amino Acid Sequence↗
PubMed1978

Sequence variation among heavy chains from inulin-binding myeloma proteins.

The entire sequences of the variable region of four heavy chains from BALB/c inulin-binding myeloma proteins have been determined. Among the four proteins there are six amino acid differences, all of which occur in the framework portion of the variable region. All of the six amino acid substitutions can be explained by single base mutations at the DNA level. The pattern of diversity in these proteins is compared to a previously reported group of heavy chains from phosphorylcholine-binding myeloma proteins. Unlike the phosphorylcholine-binding proteins, which (with the exception of two that are identical) have size and sequence differences in their complementarity regions, the inulin-binding heavy chains all have identical complementarity regions with H3 being extremely short. The pattern of variation observed in the anti-inulin heavy chains appears to be most easily explained by a somatic mutation mechanism. However, because none of the substitutions occur in complementarity-determining regions, they presumably would have no selective advantage and would not alter binding specificity. These proteins have further been shown to have crossreacting antigenic determinants (idiotypes). Five of the six sequence differences observed occur at positions that are internal in the molecule and thus presumably would not account for the idiotypic differences. These results suggest that most of the observed idiotypic crossreactivities will be due to differences in the light chains of the anti-inulin proteins.

Amino Acid Sequence↗
PubMed1977

Heavy-chain variable-region sequence from an inulin-binding myeloma protein.

The entire variable-region sequence of the heavy chain from ABE-47N, a BALB/c inulin-binding myeloma protein, has been determined. This protein is unusual in that the third complementarity region (H3) is extremely short, consisting of at the most three and probably only one amino acid. A comparison of the heavy-chain hypervariable regions from mouse, human, and rabbit proteins shows that the variability in length of H3 is greater than that seen in the first or second hypervariable regions. This variability in H3 length suggests a specialized function for this region.

Amino Acid Sequence↗
PubMed1977

Murine plasma cells secreting more than one class of immunoglobulin heavy chain. IV. Sequence differences between kappa-chains of SAMM 368 IgG2b and IgA.

The amino terminal sequences of the kappa light chains from SAMM 368 IgG2b and IgA have been determined. The two chains belong to different kappa subgroups as indicated by 10 amino acid differences among the first 23 residues. An additional three residue differences were demonstrated in the sequence including the first complementarity region. These results indicate that single cells of SAMM 368 produce two unique kappa-chains.

Amino Acid Sequence↗
PubMed1976

Expression of equivalent clonotypes in BALB/c and A/J mice after immunization with phosphorylcholine.

Analysis of A/J antibody to phosphorylcholine (PC) revealed a striking degree of similarity to PC-binding myeloma proteins of BALB/c origin. By quantitative idiotypic analysis A/J anti-PC antibody was composed to antibodies bearing binding site idiotypic determinants indistinguishable from two different BALB/c myeloma proteins, T15 and M511. Idiotypic determinants of three other PC-binding proteins, W3207, M167, and M603 were not detected. Isoelectric focusing of the light chains verified the presence of antibodies similar to T15 and M511 and indicated the presence of a third antibody whose light chains had a pI identical to that of M603. When the sequence of A/J heavy chains were compared to the heavy chains of T15, M511, and M603, both the framework and first complementarity regions were identical in all cases. Sequences analysis of the light chains through part of the first complementarity region revealed three chains, one similar to each of the myeloma proteins T15, M603, and M167-M511. The latter two sequences differ by only a single amino acid (a single base substitution) in the first 23 residues, suggesting that the two light chains may be very similar if not identical. Thus, BALB/c and A/J mice which differ genetically at multiple loci including the heavy chain allotype complex locus show a remarkable preservation of their anti-PC antibodies. These results indicate that the genes encoding these antibodies are contained in the germ line.

Amino Acid Sequence↗
PubMed1976

Size differences among immunoglobulin heavy chains from phosphorylcholine-binding proteins.

The entire sequences of the heavy chain variable regions of M167 and TEPC 15 (phosphorylcholine-binding myeloma proteins of BALB/c origin) have been determined. These sequences are compared with the phosphorylcholine-binding protein M603. T15 differs from M603 at four positions, all of which are located in antigen-binding complementarity regions. M167, in addition to having differences in the complementarity regions, also has five substitutions in the conserved framework portion of the variable region when compared to T15 and M603. Each of the three proteins has a different length in the third complementarity region. It is unlikely that complementarity regions of different lengths associated with similar framework regions could be generated by proposed mechanisms of somatic mutation which are generally limited to point mutations. It appears more likely that these products are directly encoded by different structural germ line genes.

Amino Acid Sequence↗
PubMed1976

Localized deletion-insertion mutations: a major factor in the evolution of immunoglobulin structural variability.

New and previously published amino acid sequences of mouse Vk chains completed to Trp35 were examined. This sequence includes a framework loop from NH2 to Cys23 and an antigen contacting complementarity region L1 which lies between Cys23 and Trp35. There are many different sequence variations from NH2 to Cys23 among BALB/c mouse Vk chains indicating multigenic control. Coordinately there is a remarkable structural heterogeneity of L1 size and sequence. Thus far chains with the same NH2-Cys23 sequence have similar sized L1, which suggests a close relationship between L1 size and characteristic NH2-Cys23 sequence. In this multigenic system selection for chains with size variations in L1 appears to have occurred. The mutational process that inserts or deletes triplets in this region is thought to be exploited in the derivation of new useful structural genes.

Amino Acid Sequence↗
PubMed1975

An IgM Waldenström with specificity against phosphorylcholine.

Anti-phosphorylcholine specificity has recently been shown to occur with relatively high incidence among IgA myeloma proteins secreted by oil-induced plasma cell tumors in the BALB/c strain of mice. A similar screening of human myeloma sera indicates that in man activity for phosphorylcholine is very rare. Among 904 human sera containing IgG, IgA, and IgM M-components only one reacted with phosphorylcholine-containing antigens. This serum was obtained from a patient with macroglobulinemia Waldenström. The active homogeneous protein could be isolated by affinity chromatography using a Sepharose-phosphorylcholine immunoadsorbent. It was an IgM immunoglobulin; the light chains were of the kappa type. The association constant for the reaction with phosphorylcholine was homogeneous and equalled 6.4 times 10-4 l. mol-1 at 25 degrees and 8.1 times 10-4 l. mol-1 at 2 degrees, indicating that the binding reaction is exothermic. The valences of the pentamer IgM, the 7S IgM subunit produced by reduction with cysteine, and the Fab fragment obtained by cleavage with papain were 10, 2, and 1, respectively. By all criteria available for antibody-like binding such as high specificity, restriction of the binding sites to the Fab part of the molecule and correct stoichiometry this IgM exhibits the fundamental characteristics associated with conventionally induced antibodies.

Aged↗
PubMed1975

Structural, functional, and idiotypic characteristics of a phosphorylcholine-binding IgA myeloma protein of C57BL/ka allotype.

An IgA phosphorylcholine (PC)-binding myeloma protein with IgCH allotypic determinants different from those of BALB/c mice is characterized. The myeloma, CBPC 2, was induced in the CB-20 strain of mice which is congenic to BALB/c but differs from it by carrying the A15 allotypic determinant of C57BL/ka mice. Sequence analysis of the CBPC 2 light chain through the first hypervariable region, as well as isoelectric point analysis, show that this chain is indistinguishable from that of T15, a PC-binding myeloma protein of BALB/c origin. The heavy chains of CBPC 2 and T15 differ by only two amino acids (positions 14 and 16) through the first hypervariable region. As measured by inhibition of precipitation, both CBPC 2 and T15 have the same specificity for PC, glycerophosphorylcholine, acetylcholine, and choline. In addition, CBPC 2 possesses the binding site-associated idiotypic determinant which is present on T15. However, like normal or induced C57BL/6 anti-PC antibody, it does not possess the nonbinding site idiotypic determinant.

Amino Acid Sequence↗