Recovery of binding activity in reconstituted mouse myeloma proteins

Bridges, Sandra; Little, J R

doi:10.1021/bi00789a016

Cited by 78 publications

(31 citation statements)

References 8 publications

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“…The sequence of Till differs from those of M63 and A22 (Vx-21B) by [6][7] residues, from those of M321 and T124 (VK-21C) by [6][7] residues, from that of M7Oby 22 residues, and from that of C101 by 16 The sequence of CB11 differs from the VK-21A, VK-21B, and VK-21C isotype sequences by [18][19] residues. Approximately half of the differences occur within the hypervariable regions Many of the framework region differences occur in the segment between L2 and L3 (Fig.…”

Section: Resultsmentioning

confidence: 94%

Mechanisms of antibody diversity: Multiple genes encode structurally related mouse κ variable regions

McKean¹,

Bell²,

Potter³

1978

Proc. Natl. Acad. Sci. U.S.A.

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The complete amino acid sequences of the variable regions of three mouse Vx-21 kappa chains (A22, Till, and CB10) and one partial sequence (B32) have been determined and are compared to four previously reported variable regions. These eight K variable region sequences have, with the exception of an amide difference at residue 1, identical amino-terminal 23-residue sequences, all are of the same length, and all have extensive amino acid sequence homology throughout the variable region. When these eight variable regions are grouped by sequence homology, five different groups (Vx-21A, B, C, D, and E) are present whose members share common sets of amino acids within a group. The origins of the genetic basis of antibody diversity have been approached through amino acid sequence analyses of immunoglobulin (Ig) variable regions of human, mouse, and rabbit K (VK), X (VX), and heavy chains (VH) (1), and more recently by nucleic acid hybridization (2-4). Amino acid sequences of immunoglobulin variable (V) regions derived from a single inbred strain of mice (e.g., BALB/c) most directly reflect the Ig-producing genetic potential of the individual. In such a system the role of genetic polymorphisms is reduced to a minimum and many of the differences in Ig primary structures can be related to the genetic mechanisms of Ig diversity.With the availability of several three-dimensional models of Ig V regions that have been determined by x-ray crystallography (5, 6) it has been possible to determine the functional components of the primary sequences of heavy and light chains.The framework segments of the V region sequences determine the conformation of the variable domain and interactions with other domains. Amino acid sequence differences in the framework segments of the V region are thought to be permissive changes that apparently do not alter the ability of the polypeptide to fold into a functional domain (7). The chains fold so that the complementarity-determining regions [also referred to as hypervariable regions (8) Mouse VK sequences that differ from all other mouse VK sequences by three amino acids within the amino-terminal 23 residues have been tentatively defined as an isotype. Such sequence differences within the amino-terminal 23-residue sequences are predictive of extensive sequence differences throughout the V regions whose complete sequences have been determined (10,13,17). By this criterion, 26 VK isotypes (VK-1 to VK-26) have been distinguished among 63 available sequences (16). It is expected that the criteria used to distinguish isotypes from the primary amino acid sequence data will become more refined as additional complete VK sequences become available and enable us to determine what genetic mechanisms are involved in generating antibody diversity.This involves the sequence analysis of eight V regions that have been assigned to the VK-21 isotype (16). In 1973 we (13) compared the sequence diversity in a group of four of these proteins. With the exception of an amide difference at residue 1, the four ...

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Section: Resultsmentioning

confidence: 94%

Mechanisms of antibody diversity: Multiple genes encode structurally related mouse κ variable regions

McKean¹,

Bell²,

Potter³

1978

Proc. Natl. Acad. Sci. U.S.A.

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“…Heavy and light chains of myeloma proteins and anti-PC antibodies were isolated after partial reduction and alkylation by chromatography on Sephadex G100 columns equilibrated with 1 M propionic acid, 4.5 M urea (25) . The ascending and descending portions of the heavy and light chain peaks, respectively, were used .…”

Section: Methodsmentioning

confidence: 99%

Clonal nature of the immune response to phosphorylcholine (PC). V. Cross-idiotypic specificity among heavy chains of murine anti-PC antibodies and PC-binding myeloma proteins.

Claflin¹,

Davie²

1975

The Journal of Experimental Medicine

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Williams et al.(1) in a study of monoclonal IgM cold agglutinins noted that antisera to different cold agglutinins possessed specificity not only for antigenic determinants unique to the individual proteins but also for determinants that were shared by cold agglutinins from a number of individuals . This phenomenon of shared or cross-idiotypy among immunoglobulins with similar specificity also has been demonstrated for human proteins with anti-7-globulin activity (2) and among rabbit (3, 4) and mouse (5) antibodies to streptococcal group carbohydrates . In most instances the proteins exhibiting cross-idiotypic reactions had similar, but not necessarily identical, combining sites, and the idiotypic determinant shared by them was associated with the combining site (2, 5-8) . Support for the structural similarity of antibodies with similar specificity and shared idiotypy has been provided by recent findings showing that IgM cold agglutinins, anti-y-globulins (9, 11), or rabbit antibodies to streptococcal group C carbohydrate (7) have strikingly similar heavy and/or light chains .Previous studies from our laboratory have demonstrated a remarkable similarity in the combining sites of mouse antibodies to phosphorylcholine (PC)' regardless of the genetic background of the mice. The antibodies from the mouse strains tested possessed indistinguishable binding specificities for a number of choline analogues (12) and had identical binding-site antigenic determinants (13) . This suggests a striking conservation of the binding site region of mouse anti-PC antibodies, even though differences have been shown to exist in other portions of the variable region (14-16) . In contrast, it was shown (17) that three PC-binding myeloma proteins that differed in combining specificity (18), as well as in idiotypes (19) and in light chains (17), have heavy chain sequences which are virtually identical through the first hypervariable region . These findings *

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“…The heavy chains and light chains of antibody 2B6 were separated by reduction of the disulfide bonds of the antibody with dithiothreitol. 17 To an aqueous solution of antibody 2B6 (6.6 © 10 ¹6 M), ethylenediaminetetraacetic acid tetrasodium salt (EDTA-4Na, 1.0 mM) was dissolved. Dithiothreitol (0.1 M) in 0.5 mL of tris buffer (pH 7.0) was added to the solution.…”

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confidence: 99%

Photoinduced Hydrogen-Evolution System with an Antibody–Porphyrin Complex as a Photosensitizer

Yamaguchi¹,

Onji²,

Ohara³

et al. 2009

Bulletin of the Chemical Society of Japan

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A complex between a monoclonal antibody for porphyrin and zincporphyrin was utilized to construct an energy conversion system. Monoclonal antibody 2B6 bound meso-tetrakis(4-carboxyphenyl)porphyrin zinc complex (ZnTCPP) with a dissociation constant of 2.1 © 10 ¹8 M. Upon binding the antibody, the lifetime of the excited triplet state of ZnTCPP increased from 0.5 to 1.2 ms. A stable cationic radical of viologen was obtained by irradiating the solution containing the complex of 2B6 with ZnTCPP, methyl viologen (MV 2+ ), and ethylenediaminetetraacetic acid tetrasodium salt (EDTA-4Na) with light. When colloidal platinum was added as a catalyst, photoinduced hydrogen production was observed upon continuous irradiation of visible light. The estimated turnover number of photoinduced hydrogen evolution was 5.0 © 10 ¹3 s ¹1. The catalytic activity of the 2B6ZnTCPP complex on the hydrogen evolution was compared with that of ZnTCPP alone and the complexes of ZnTCPP with fragments of antibody 2B6 (2B6-H and 2B6-L). The heavy chain of antibody 2B6 mainly contributed to the complex formation with ZnTCPP and the resultant hydrogen production, and the whole antibodyZnTCPP complex led to the efficient hydrogen production.Recently, the design of new energy sources has received much attention because the exhaustion of fossil fuels has become a pressing issue. Furthermore, combustion of fossil fuels causes emission of CO 2 , which has resulted in global warming. Hence, solar energy has become popular due to its limitlessness. Moreover energy derived from hydrogen has received a lot of attention as a clean energy source. Currently, there are many attempts to create energy conversion systems, which convert solar energy into chemical energy.1 On the other hand plants and light harvesting bacteria convert solar energy into chemical energy with high efficiency. 2In the field of artificial energy conversion, hydrogen production systems by photoelectrolysis of water constructed using TiO 2 electrodes have been developed. 3 However, TiO 2 absorbs only UV light, and it cannot use the major part of sunlight. To overcome this problem, dye-sensitized hydrogen-evolution systems have been developed. 4 It is advisable to mimic the energy conversion systems in nature, and the chromophores of in vivo photosynthetic reaction centers are fixed by the protein environment. 5 The distances between electron donors and electron acceptors are noncovalently held under optimum conditions. Recently, non-covalently assembled donoracceptor arrays have been constructed utilizing hydrogen bonding, 6 metal coordination, 7 apoproteins, 8 electrostatic interactions, 9 and supramolecular formations. 10To construct a photoinduced hydrogen production system, a suitable catalyst is necessary to reduce a proton by an electron, which can be obtained by charge separation. In many cases, colloidal platinum has been used as a catalyst for proton reduction.11 To construct an artificial charge-separation system with non-covalently linked electron donors and acceptors, it is fa...

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Recovery of binding activity in reconstituted mouse myeloma proteins

Cited by 78 publications

References 8 publications

Mechanisms of antibody diversity: Multiple genes encode structurally related mouse κ variable regions

Mechanisms of antibody diversity: Multiple genes encode structurally related mouse κ variable regions

Clonal nature of the immune response to phosphorylcholine (PC). V. Cross-idiotypic specificity among heavy chains of murine anti-PC antibodies and PC-binding myeloma proteins.

Photoinduced Hydrogen-Evolution System with an Antibody–Porphyrin Complex as a Photosensitizer

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