Structure of the heavy chain from strain 13 guinea pig immunoglobulin G1: isolation of cyanogen bromide fragments

Guinea pigs immunized with 2,4-dinitrophenyl-guinea pig albumin (DNP-GPA) possess lymphocytes which specifically bind sufficient DNP-GPA-125I to their surface to be detected by radioautography. These lymphocytes are present in the draining lymph nodes in a frequency of ∼50/1000 lymphocytes in animals immunized 2–4 wk earlier with DNP-GPA in complete Freund's adjuvant. Nonimmunized animals have ∼0.4 DNP-GPA antigen-binding cells (ABC) per 1000 lymphocytes. An increase in the frequency of DNP-GPA ABC in peripheral blood is detectable by 5 days after immunization, which is before the time that serum anti-DNP antibody is measurable. The receptors of these ABC are hapten specific in that free ϵ-DNP-L-lysine, at low concentration, inhibits the binding of DNP-GPA-125I; DNP bovine serum alumbin (DNP-BSA) is equivalent to DNP-GPA in the inhibition of binding of DNP-GPA-125I to ABC; and both DNP-GPA agarose beads and DNP-BSA agarose beads specifically adsorb DNP-GPA-125I ABC. Anti-immunoglobulin antisera, particularly anti-γ2 sera, inhibit the binding of DNP-GPA-125I to these cells implying that the receptors are immunoglobulin, primarily of the γ2 heavy chain class. DNP-GPA-125I ABC appear to represent precursors of antibody-secreting cells and have specificity characteristics which are very different from cells, of similarly immunized guinea pigs, which mediate a cellular immune response to DNP-GPA.

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The protein-synthesizing activity of ribosomes isolated from the mammary gland of lactating and pregnant guinea pigs

Biochemical Journal ◽

10.1042/bj1230865 ◽

1971 ◽

Vol 123 (5) ◽

pp. 865-874 ◽

Cited By ~ 18

Author(s):

E. Fairhurst ◽

Diana McIlreavy ◽

P. N. Campbell

Keyword(s):

Protein Synthesis ◽

Mammary Gland ◽

Guinea Pig ◽

Guinea Pigs ◽

Cyanogen Bromide ◽

Mammary Glands

1. Polyribosome preparations were made from the deoxycholate-treated post-nuclear fractions obtained by the disruption of mammary glands from lactating and pregnant guinea pigs. 2. A high proportion of large polyribosomes was obtained from the glands of lactating animals whereas mainly small polyribosomes were obtained from the glands of pregnant animals. The isolated preparations incorporated [14C]phenylalanine into protein. The polyribosomes from the glands of pregnant animals were less active than those from the glands of lactating animals but the activity of the former was stimulated more by poly(U) than was the latter. 3. The ribosomes from mammary gland could be dissociated into subunits after incubation, under conditions necessary for protein synthesis, in the presence of puromycin. The subunits could be recombined to give a preparation that actively polymerized [14C]phenylalanine in the presence of poly(U). The subunits from guinea-pig mammary gland could be combined with subunits from liver of either guinea pig or rat. Hybrid ribosomes were also formed from subunits derived from glands of pregnant and lactating animals. The hybrids were as active as were the ribosomes formed by reassociation of subunits from the same tissue, suggesting that in this respect the ribosomes from pregnant animals were not defective. 4. Polyribosomes from mammary glands of lactating animals when incubated with cell sap from the same source were tested for their ability to synthesize α-lactalbumin. The polyribosomes were incubated in the presence of [3H]leucine and α-lactalbumin was isolated from the supernatant. The protein was finally treated with cyanogen bromide and the C-terminal and N-terminal fragments were separated and their radioactivity was determined. Both fragments were radioactive consistent with the synthesis of α-lactalbumin. 5. The results are discussed in relation to protein synthesis in the mammary gland after parturition.

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The Order of Cyanogen-Bromide Peptides and Location of Carbohydrate in the alpha2 Chain of Guinea-Pig Skin Collagen

European Journal of Biochemistry ◽

10.1111/j.1432-1033.1975.tb02237.x ◽

1975 ◽

Vol 56 (2) ◽

pp. 327-333 ◽

Cited By ~ 10

Author(s):

Charles C. CLARK ◽

Peter P. FIETZEK ◽

Paul BORNSTEIN

Keyword(s):

Guinea Pig ◽

Cyanogen Bromide ◽

Pig Skin ◽

Skin Collagen

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Cyanogen bromide cleavage and partial sequence of the heavy chain of a pathological immunoglobulin G

Biochemical Journal ◽

10.1042/bj1040616 ◽

1967 ◽

Vol 104 (2) ◽

pp. 616-626 ◽

Cited By ~ 25

Author(s):

PJ Piggot ◽

EM Press

Keyword(s):

Heavy Chain ◽

Immunoglobulin G ◽

Cyanogen Bromide ◽

Partial Sequence

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Completion of the analysis of the primary structure of the variable domain of a homogeneous rabbit antibody to type III pneumococcal polysaccharide

Biochemical Journal ◽

10.1042/bj1430723 ◽

1974 ◽

Vol 143 (3) ◽

pp. 723-732 ◽

Cited By ~ 14

Author(s):

Jean-Claude Jaton

Keyword(s):

Amino Acid ◽

Amino Acid Sequence ◽

Guinea Pig ◽

Heavy Chain ◽

Variable Region ◽

Complete Sequence ◽

Rabbit Antibody ◽

Type Iii ◽

Minor Variant ◽

V Region

The amino acid sequence between residues 70 and 116 of the V (variable) region of the H (heavy) chain derived from rabbit antibody BS-5, specific for type III pneumococcal polysaccharide, was determined. The sequence of this section of the H chain which includes the hypervariable residues 94 to about 112 was unique, although minor variant sequences present in the H chain preparation would not have been detected by the techniques used in this work. Taken together with the known sequences of the N-terminal 69 residues of H chain BS-5 (Jaton & Braun, 1972) and of the V region of the light chain (Jaton, 1974b), the data establish the complete sequence of the V domain of a rabbit immunoglobulin G. The V region of H chain BS-5 is compared with the basic sequences of the three human V region subgroups known to date, with one mouse H chain, and with guinea-pig pooled H chains. Even though chains from guinea pig and mouse clearly belong to the subgroup III of variability (VHIII), rabbit H chain BS-5 (allotypic variant a1) appears more closely related to the subgroup VHII than to the subgroups VHIII or VHI. The homology between VL and VH regions of antibody BS-5 (28%) is not greater than that observed between the VH region of antibody BS-5 and the VL regions of different rabbit antibodies.

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