RESUMO
We herein report a case of a combined crystalline light chain tubulopathy, podocytopathy, histiocytosis, and cast nephropathy in a patient with monoclonal gammopathy of renal significance (MGRS). A 66-year-old female with impaired renal function was referred to our department. Despite intravenous fluid resuscitation, the kidney function worsened progressively; thus, a kidney biopsy was performed. The kidney biopsy revealed light chain proximal tubulopathy (LCPT) with crystals, light chain crystal podocytopathy (LCCP), crystal-storing histiocytosis (CSH), and light chain cast nephropathy (LCCN). Of note, LCCP and CSH were diagnosed via electron microscopy. Serum and urine immunoelectrophoresis (IEP) revealed the presence of monoclonal Bence-Jones protein and free κ light chains. Bone marrow aspiration showed < 10% plasma cell proliferation. Thus, we had encountered a rare case in which a variety of kidney lesions were combined with MGRS. Most of the LCPT, LCCP, and CSH cases show monoclonal IgG κ, while our case showed Bence-Jones protein κ.
Assuntos
Proteína de Bence Jones/isolamento & purificação , Histiocitose/complicações , Nefropatias/diagnóstico , Idoso , Feminino , Humanos , Cadeias kappa de Imunoglobulina , Nefropatias/etiologia , Túbulos Renais Proximais/patologia , Microscopia Imunoeletrônica , Podócitos/patologiaRESUMO
Deposition of immunoglobulin light chains is a result of clonal proliferation of monoclonal plasma cells that secrete free immunoglobulin light chains, also called Bence Jones proteins (BJP). These BJP are present in circulation in large amounts and excreted in urine in various light chain diseases such as light chain amyloidosis (AL), light chain deposition disease (LCDD) and multiple myeloma (MM). BJP from patients with AL, LCDD and MM were purified from their urine and studies were performed to determine their secondary structure, thermodynamic stability and aggregate formation kinetics. Our results show that LCDD and MM proteins have the lowest free energy of folding while all proteins show similar melting temperatures. Incubation of the BJP at their melting temperature produced morphologically different aggregates: amyloid fibrils from the AL proteins, amorphous aggregates from the LCDD proteins and large spherical species from the MM proteins. The aggregates formed under in vitro conditions suggested that the various proteins derived from patients with different light chain diseases might follow different aggregation pathways.
Assuntos
Amiloidose , Proteína de Bence Jones/química , Mieloma Múltiplo , Dobramento de Proteína , Amiloidose/metabolismo , Proteína de Bence Jones/isolamento & purificação , Proteína de Bence Jones/metabolismo , Humanos , Cadeias Leves de Imunoglobulina , Mieloma Múltiplo/metabolismo , Estrutura Secundária de Proteína , TermodinâmicaRESUMO
Up to 3% of adults over 50 years of age show a monoclonal peak values in blood or urine. Findings and prognosis will be distinct in view of the nature of this factor. In B-cell neoplasias (multiple myeloma, Waldeström macroglobulinaemia, chronic myeloid leukaemia and non-Hodgkin lymphoma) the clinical pattern is dominated by the systemic effects produced by the expansion of the malign clone; the monoclonal protein may result in hyperviscosity syndrome or renal damage. On the other hand, there are other less frequent processes called diseases associated to monoclonal components, where the main clinical manifestations and prognosis depend of the biological effects of the monoclonal protein. With reference to this last group, which is the objective of this revision, no bone lesions, anaemia or a greater tendency to infections usually occur when compared with the first group. Even so, there are some cases of interposition between both groups: for instance, type IgM immunoglobulin present in Waldeström macroglobulinaemia may have cold agglutinin activity, and in the case of multiple myeloma, the clone may secrete amyloidogenic light chains (AU)
Assuntos
Humanos , Masculino , Feminino , Proteína de Bence Jones , Proteína de Bence Jones/imunologia , Proteína de Bence Jones/isolamento & purificação , Paraproteinemias/imunologia , Paraproteinemias/diagnóstico , Paraproteinemias/metabolismo , PrognósticoRESUMO
Four crystal forms have been grown and characterized by X-ray diffraction of a Bence-Jones protein collected from the urine of a multiple myeloma patient more than 40 years ago. Closely related tetragonal and orthorhombic forms belonging to space groups P4(3)2(1)2 and P2(1)2(1)2(1), with unit-cell parameters a = b = 68.7, c = 182.1 and a = 67.7, b = 69.4, c = 87.3 A, diffract to 1.5 and 1.9 A, respectively. Two closely related trigonal forms, both belonging to space group P3(1)21 with unit-cell parameters a = b = 154.3 A but differing by a doubling of the c axis, one 46.9 A and the other 94.0 A, diffract to 2.9 and 2.6 A resolution, respectively. The trigonal crystal of short c-axis length shows a positive indication of twinning. The trigonal crystal of longer c axis, which appeared only after eight months of incubation at room temperature, is likely to be composed of proteolytically degraded molecules and unlike the other crystal forms contains two entire Bence-Jones dimers in the asymmetric unit. This latter crystal form may shed some light on the formation of fibrils common to certain storage diseases.
Assuntos
Proteína de Bence Jones/química , Mieloma Múltiplo/sangue , Adulto , Proteína de Bence Jones/isolamento & purificação , Cristalização , Liofilização , Humanos , Pessoa de Meia-Idade , Conformação Proteica , Difração de Raios XAssuntos
Troca Plasmática , Plasmaferese , Macroglobulinemia de Waldenstrom/terapia , Anticorpos Monoclonais/sangue , Anticorpos Monoclonais/isolamento & purificação , Proteína de Bence Jones/isolamento & purificação , Viscosidade Sanguínea , Humanos , Imunoglobulina A/isolamento & purificação , Imunoglobulina G/isolamento & purificação , Imunoglobulina M/sangue , Imunoglobulina M/isolamento & purificação , Neoplasias/complicações , Troca Plasmática/métodos , Plasmaferese/métodos , Macroglobulinemia de Waldenstrom/diagnóstico , Macroglobulinemia de Waldenstrom/etiologiaRESUMO
Crystals of a human (Sea) Bence-Jones dimer were produced in a capillary by vapor diffusion under microgravity conditions in the 9 day US Space Shuttle Mission STS-95. In comparison to ground-based experiments, nucleation was facile and spontaneous in space. Appearance of a very large (8 x 1.6 x 1.0 mm) crystal in a short time period is a strong endorsement for the use of microgravity to produce crystals sufficiently large for neutron diffraction studies. The Sea dimer crystallized in the orthorhombic space group P2(1)2(1)2(1), with a = 48.9 A, b = 85.2 A, and c = 114.0 A. The crystals grown in microgravity exhibited significantly lower mosaicities than those of ground-based crystals and the X-ray diffraction data had a lower overall B factor. Three-dimensional structures determined by X-ray analysis at two temperatures (100 and 293 K) were indistinguishable from those obtained from ground-based crystals. However, both the crystallographic R factor and the free R factor were slightly lower in the models derived from crystals produced in microgravity. The major difference between the two crystal growth systems is a lack of convection and sedimentation in a microgravity environment. This environment resulted in the growth of much larger, higher-quality crystals of the Sea Bence-Jones protein. Structurally, heretofore unrecognized grooves on the external surfaces of the Sea and other immunoglobulin-derived fragments are regular features and may offer supplementary binding regions for super antigens and other elongated ligands in the bloodstream and perivascular tissues.
Assuntos
Proteína de Bence Jones/química , Voo Espacial , Ausência de Peso , Proteína de Bence Jones/isolamento & purificação , Cristalização , Cristalografia por Raios X , Humanos , Modelos Moleculares , Mieloma Múltiplo/urina , Difração de Nêutrons , Conformação Proteica , Água/químicaRESUMO
We report here a lupus anticoagulant (LA)-like activity observed in a 45-year-old man with Bence-Jones protein (BJP) lambda-type multiple myeloma. This patient showed no clinical symptoms of thrombosis or bleeding diathesis. Laboratory examination on admission showed mild anemia, prolongation of activated partial thromboplastin time (APTT) (APTT, 56.2 seconds; control, 29.1 seconds), normal prothrombin time, normal thrombin time, and massive proteinuria (2.3 g/d). The mix test with normal plasma showed the presence of circulating anticoagulant. Based on the assumption that the lambda-type BJP may have been responsible for the prolongation of APTT, we purified the BJP from the patient's urine using column works. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting showed that the purified protein was a 48-kd homodimer of immunoglobulin lambda-chains. Addition of the purified dimeric lambda-type BJP to the normal plasma prolonged both APTT and dilute Russell's viper venom time (DRVVT) in a dose-dependent manner, and the negatively charged phospholipid-dependent prothrombinase activity was significantly inhibited in the presence of this protein. Furthermore, both the prolongation of DRVVT and the inhibition of the prothrombinase activity were almost completely abrogated under the condition of high ionic strength. These findings collectively suggest that the dimeric lambda-type BJP showed LA-like activity via the mechanism of ionic charge.
Assuntos
Proteína de Bence Jones/farmacologia , Inibidor de Coagulação do Lúpus , Mieloma Múltiplo/sangue , Proteína de Bence Jones/isolamento & purificação , Proteína de Bence Jones/urina , Coagulação Sanguínea/efeitos dos fármacos , Dimerização , Humanos , Cadeias lambda de Imunoglobulina/isolamento & purificação , Cadeias lambda de Imunoglobulina/farmacologia , Cadeias lambda de Imunoglobulina/urina , Masculino , Pessoa de Meia-Idade , Tempo de Tromboplastina ParcialRESUMO
Patients with systemic amyloidosis associated with multiple myeloma (AL-amyloidosis) exhibit immunoglobulin light chains and fragments which have been identified as amyloid protein. Since a relatively small proportion of patients with multiple myeloma develop AL-amyloidosis, comparison of the amino acid sequence of the amyloidogenic and non-amyloidogenic immunoglobulin light chains and the structural characterization of the amyloid proteins are required to understand the relationship between structure and amyloidogenicity. We determined the primary structure of a kappa I-type Bence Jones protein obtained from a patient (FUR) who had systemic AL-amyloidosis associated with multiple myeloma. We identified eight amino acid replacements unique to this patient among the amyloidogenic kappa I-light chains, and which are also rare among the known kappa type light chains of humans. Three of these substitutions were within the framework regions and may act to destabilize the structure to promote a putative amyloidogenic conformation. In contrast to light chain fragments in the urine, which were processed in the variable region, mass spectrometric analysis of the fibril proteins isolated from lingual amyloid deposits in this patient, revealed that they were all truncated within the constant region and corresponded to residues 1-125, 1-144, and 1-210. Inspection of the predicted three-dimensional model of this protein suggested that these fragments may be generated by a protease specific for the N-terminal sides of basic amino acids. These findings suggest that amino acid substitutions at highly conserved residues may convert non-amyloidogenic to amyloidogenic immunoglobulin light chain proteins.
Assuntos
Proteína de Bence Jones/química , Idoso , Sequência de Aminoácidos , Amiloidose/complicações , Amiloidose/metabolismo , Proteína de Bence Jones/isolamento & purificação , Proteína de Bence Jones/urina , Evolução Fatal , Humanos , Cadeias Leves de Imunoglobulina/análise , Cadeias kappa de Imunoglobulina/análise , Masculino , Modelos Moleculares , Dados de Sequência Molecular , Peso Molecular , Mieloma Múltiplo/complicações , Conformação Proteica , Língua/patologiaRESUMO
Immunochemical analysis of lambda type Bence Jones protein (BJP: designated as Suzuki-BJP) and IgG-lambda type M-protein (designated as Miki-IgG), lambda type BJP (designated as Miki-BJP) which showed non-specific binding with lactate dehydrogenase (LD, EC 1.1.1.27) was carried out in two cases. When the purified LD mixed with NADH was eluted through the CNBr-Sepharose 4B coupled to Suzuki-BJP or Miki-IgG, the affinity with these adsorbents was not demonstrated. The amino acid residue of the N-terminal in the Suzuki-BJP and lambda chain of the Miki-IgG was determined to be tyrosine by primary structure analysis, on the other hand, alanine was detected in the gamma chain of the Miki-IgG that did not have LD binding ability. By counter affinity electrophoresis, it was shown that LD bound to a synthetic peptide consisting of 15 amino acid residues of N-terminal which had the same beta-sheet structure as the Suzuki-BJP. It seems probable that LD combines with BJP (or IgG) molecule at the NAD+ binding site producing a three-dimensional structure similar to NAD+.
Assuntos
Proteína de Bence Jones/metabolismo , Imunoglobulina G/metabolismo , L-Lactato Desidrogenase/metabolismo , Idoso , Idoso de 80 Anos ou mais , Sequência de Aminoácidos , Proteína de Bence Jones/química , Proteína de Bence Jones/isolamento & purificação , Cromatografia de Afinidade , Brometo de Cianogênio , Feminino , Humanos , Imunoglobulina G/química , Imunoglobulina G/isolamento & purificação , Imuno-Histoquímica , Isoenzimas , Masculino , Pessoa de Meia-Idade , Dados de Sequência Molecular , Estrutura Secundária de Proteína , SefaroseRESUMO
Cardiomyopathy due to monoclonal light chain deposits is a complication of plasma cell disorders. The deposits may be either fibrillar as in light chain amyloid or nonfibrillar as in light chain deposition disease. The reasons for these structural differences are still unknown. We characterized the myocardial deposits by immunohistochemical examination of sections and extraction and biochemical analysis of the tissue deposits in a patient (MCM) who died of myeloma and systemic light chain deposition disease. Amino acid sequence analysis of the extracted nonfibrillar MCM kappa-light chain reveals that it belongs to the L12a germline subset of the kappa(I) protein and contains five distinctive amino acid substitutions (three in the framework region III and two in the complementarity-determining region III) that have not been reported previously in the same positions in other kappa(I) light chains. The theoretically determined isoelectric point (pI 8.21) of the MCM light chain is high compared with the low isoelectric point of other Bence Jones proteins from subjects without light chain deposition disease. The diffuse binding to basement membranes and the high isoelectric point of the MCM kappa-light chain suggest electrostatic interaction as a possible mechanism of tissue deposition. The spatial locations of the five distinctive residues and a sixth rare substitution of the MCM protein modeled on the backbone structure of REI, a kappa(I)-soluble Bence Jones light chain of known three-dimensional structure, may be responsible for protein destabilization, partial unfolding, and aggregation leading to tissue deposition.
Assuntos
Aminoácidos/análise , Cardiomiopatias/patologia , Cadeias Leves de Imunoglobulina/análise , Cadeias Leves de Imunoglobulina/química , Miocárdio/química , Miocárdio/patologia , Adulto , Sequência de Aminoácidos , Proteína de Bence Jones/análise , Proteína de Bence Jones/química , Proteína de Bence Jones/isolamento & purificação , Cardiomiopatias/etiologia , Cardiomiopatias/imunologia , Humanos , Hipergamaglobulinemia/complicações , Hipergamaglobulinemia/imunologia , Hipergamaglobulinemia/patologia , Cadeias Leves de Imunoglobulina/isolamento & purificação , Cadeias kappa de Imunoglobulina/análise , Cadeias kappa de Imunoglobulina/isolamento & purificação , Cadeias kappa de Imunoglobulina/fisiologia , Imuno-Histoquímica , Ponto Isoelétrico , Masculino , Microscopia Eletrônica , Dados de Sequência Molecular , Mieloma Múltiplo/complicações , Mieloma Múltiplo/imunologia , Mieloma Múltiplo/patologia , Miocárdio/ultraestruturaRESUMO
The primary structural features that render human monoclonal light chains amyloidogenic are presently unknown. To gain further insight into the physical and biochemical factors that result in the pathologic deposition of these proteins as amyloid fibrils, we have selected for detailed study three closely homologous protein products of the light-chain variable-region single-gene family VkIV. Two of these proteins, REC and SMA, formed amyloid fibrils in vivo. The third protein, LEN, was excreted by the patient at levels of 50 g/day with no indication of amyloid deposits. Sequences of amyloidogenic proteins REC and SMA differed from the sequence of the nonpathogenic protein LEN at 14 and 8 amino acid positions, respectively, and these amino acid differences have been analyzed in terms of the three-dimensional structure of the LEN dimer. To provide a replenishable source of these human proteins, we constructed synthetic genes coding for the REC, SMA, and LEN variable domains and expressed these genes in Escherichia coli. Immunochemical and biophysical comparisons demonstrated that the recombinant VkIV products have tertiary structural features comparable to those of the patient-derived proteins. This well-defined set of three clinically characterized human kIV light chains, together with the capability to produce these kIV proteins recombinantly, provide a system for biophysical and structural comparisons of two different amyloidogenic light-chain proteins and a nonamyloidogenic protein of the same subgroup. This work lays the foundation for future investigations of the structural basis of light-chain amyloidogenicity.
Assuntos
Amiloide/imunologia , Amiloidose/imunologia , Proteína de Bence Jones/imunologia , Região Variável de Imunoglobulina/imunologia , Cadeias kappa de Imunoglobulina/imunologia , Sequência de Aminoácidos , Sequência de Bases , Proteína de Bence Jones/isolamento & purificação , Genes Sintéticos , Humanos , Região Variável de Imunoglobulina/genética , Cadeias kappa de Imunoglobulina/genética , Modelos Moleculares , Dados de Sequência Molecular , Conformação Proteica , Desnaturação Proteica , Proteínas Recombinantes de Fusão/imunologia , Proteínas Recombinantes/imunologia , Homologia de Sequência de Aminoácidos , Homologia de Sequência do Ácido NucleicoRESUMO
Bence Jones proteins purified from urine of patients with multiple myeloma were found to be capable of hydrolyzing carbobenzoxy-L-valyl-glycyl-L-arginine p-nitroanilide (Chromozym TRY) and benzoyl-L-arginine p-nitroanilide (BApNA), synthetic chromogenic substrates for trypsin. The amidolytic activity obeyed classic Michaelis-Menten kinetics, exhibiting optimal activity around pH 8.4 and apparent Km of 140-730 microM and 18-27 microM for Chromozym TRY and BApNA, respectively. No activity was detected with intact IgG or Fab fragment, whereas the activity comparable to those of Bence Jones proteins was found with light chain derived from inactive IgG. Several lines of circumstantial evidence indicate that the observed activity was not due to contaminating enzyme.
Assuntos
Amidoidrolases/metabolismo , Proteína de Bence Jones/metabolismo , Amidoidrolases/isolamento & purificação , Amidoidrolases/urina , Sequência de Aminoácidos , Proteína de Bence Jones/isolamento & purificação , Proteína de Bence Jones/urina , Cromatografia de Afinidade , Cromatografia em Gel , Humanos , Cinética , Dados de Sequência Molecular , Mieloma Múltiplo/urina , Oligopeptídeos , Especificidade por Substrato , Tripsina/metabolismoRESUMO
REIv--the variable domain of an immunoglobulin x light chain--was produced by heterologous gene expression in a Gram-positive bacterium, purified to homogeneity and characterized. A host/vector combination based on secretion of Staphylococcus hyicus lipase by Staphylococcus carnosus was exploited. A gene encoding a fusion protein, composed of an aminoterminal portion of the pre-pro-peptide of S. hyicus lipase, a hexahistidine affinity tag, followed by the recognition sequence of IgA protease and REIv was constructed. Expression of the fusion gene in S. carnosus causes selective secretion and accumulation of a soluble fusion protein in the culture medium (5-10 mg/l), which can be purified from the supernatant by immobilized metal ion affinity chromatography (IMAC). REIv is released from the fusion protein with an additional threonine and proline residue at the aminoterminus (REIvTP) by site-specific cleavage with IgA protease and can be separated from the hexahistidine-tagged fusion partner and the protease by a second passage through an IMAC gel matrix. Like authentic REIv, the isolated protein (> 1 mg/l culture medium) migrates as a dimer in gel filtration chromatography and undergoes cooperative, reversible unfolding in urea. The isolated immunoglobulin REIvTP and authentic REIv have indistinguishable free energies of unfolding (approx. 26 kJ/mol, 6.3 kcal/mol).
Assuntos
Proteína de Bence Jones/biossíntese , Região Variável de Imunoglobulina/genética , Cadeias kappa de Imunoglobulina/biossíntese , Staphylococcus/metabolismo , Sequência de Aminoácidos , Sequência de Bases , Proteína de Bence Jones/análise , Proteína de Bence Jones/isolamento & purificação , Cromatografia em Gel , Escherichia coli/genética , Escherichia coli/metabolismo , Vetores Genéticos/fisiologia , Hidrólise , Cadeias kappa de Imunoglobulina/análise , Cadeias kappa de Imunoglobulina/isolamento & purificação , Dados de Sequência Molecular , Peso Molecular , Plasmídeos/genética , Plasmídeos/metabolismo , Conformação Proteica , Proteínas Recombinantes de Fusão/biossíntese , Proteínas Recombinantes de Fusão/metabolismo , Espectrometria de Fluorescência , UreiaRESUMO
Bence Jones proteins (BJPs) are the major pathogenic factor causing cast nephropathy ("myeloma kidney") by coaggregation with Tamm-Horsfall glycoprotein (THP). Understanding the interaction between these proteins is therefore important in developing treatment strategies to prevent renal failure from cast formation in multiple myeloma. We developed an enzyme-linked immunoassay to examine this phenomenon. Five different human BJPs (four kappa and one lambda immunoglobulin light chains) were used in this assay that demonstrated these proteins bound THP with different affinity. BJPs competed among themselves for binding to THP. The binding site was a peptide portion of THP since these proteins also bound deglycosylated THP. Also, one monoclonal antibody directed against a peptide segment of human THP prevented binding of THP to BJPs. By altering the conformation of THP, reducing agents decreased binding between these two proteins in concentration-dependent fashion. In turbidity studies, the monoclonal antibody that prevented binding and a reducing agent, dithiothreitol, decreased coaggregation. Deglycosylated THP did not coaggregate with BJPs. We concluded that ionic interaction between BJPs and a specific peptide binding site on THP promoted heterotypic coaggregation. The carbohydrate moiety of THP was also essential for coaggregation, perhaps by facilitating homotypic aggregation of THP.
Assuntos
Proteína de Bence Jones/metabolismo , Mucoproteínas/metabolismo , Proteína de Bence Jones/isolamento & purificação , Proteína de Bence Jones/urina , Sítios de Ligação , Ligação Competitiva , Humanos , Cadeias kappa de Imunoglobulina/metabolismo , Cadeias lambda de Imunoglobulina/metabolismo , Cinética , Masculino , Mucoproteínas/isolamento & purificação , Mucoproteínas/urina , Mieloma Múltiplo/urina , Nefelometria e Turbidimetria , Penicilamina/farmacologia , Insuficiência Renal/urina , Compostos de Sulfidrila/farmacologia , Fatores de Tempo , UromodulinaRESUMO
Motivated by the finding that the amino acid sequence of the Bence Jones protein BJP-DIA was identical to that of the main protein component of the amyloid fibrils obtained from the same patient with AL-amyloidosis, (Klafki, H.-W., Kratzin, H.-D., Pick, A.-I., Eckart, K., Karas, M. & Hilschmann, N. (1992) Biochemistry 31, 3265-3272.), we attempted to create "amyloid-like" fibrils from the Bence Jones protein in vitro, without addition of proteolytic enzymes. Reduction of BJP-DIA, solubilized in PBS, pH 7.4, overnight at 37 degrees C resulted in the formation of a precipitate which had affinity for the dye Congo red. Electron microscopy of negatively stained samples of the reduced protein revealed aggregates of linear unbranched fibrils. SDS-polyacrylamide gel electrophoresis demonstrated that the precipitate consisted almost exclusively of intact light chain molecules. This result makes it possible to deduce a molecular model of these amyloid fibrils generated in vitro.
Assuntos
Amiloide/biossíntese , Proteína de Bence Jones/química , Dissulfetos/química , Proteína de Bence Jones/isolamento & purificação , Proteína de Bence Jones/ultraestrutura , Cromatografia por Troca Iônica , Eletroforese em Gel de Poliacrilamida , Humanos , Microscopia Eletrônica , OxirreduçãoRESUMO
The complete primary structures of both the main amyloid fibril protein component (AL-DIA) and the soluble Bence Jones protein (BJP-DIA) obtained from the same patient with AL-amyloidosis are reported for the first time. The amino acid sequences were determined by automated Edman degradation following proteolytic digestion of the isolated proteins and HPLC separation of the resulting fragments and by amino-terminal sequencing after treatment with pyroglutamate aminopeptidase. Sequencing data were confirmed by amino acid analysis and plasma desorption mass spectrometry (PDMS). Molecular weights of the complete proteins were determined by laser desorption mass spectrometry. The amyloid fibril preparation contained a complete monoclonal lambda immunoglobulin light chain (subgroup 1.2) as well as different-sized fragments thereof which were identified by immunoblotting and amino-terminal sequencing following immobilization of electrophoretically-separated proteins on poly(vinylidene difluoride) (PVDF) membranes. The soluble urinary Bence Jones protein (BJP-DIA) was a dimer of monoclonal L-chains with a primary structure identical to that of the amyloid L-chain (AL-DIA) and thus represented the amyloid precursor protein.
Assuntos
Amiloide/isolamento & purificação , Amiloidose/metabolismo , Proteína de Bence Jones/isolamento & purificação , Idoso , Sequência de Aminoácidos , Amiloide/química , Proteína de Bence Jones/química , Proteína de Bence Jones/urina , Feminino , Humanos , Cadeias Leves de Imunoglobulina/isolamento & purificação , Fígado/química , Espectrometria de Massas , Dados de Sequência Molecular , Peso Molecular , Conformação ProteicaRESUMO
The authors report the clinical features and the results of immunochemical studies of patients with angioimmunoblastic lymphadenopathy, primary amyloidosis, and multiple myeloma, each of whom had in the serum and urine multiple forms of Bence Jones protein (BJP). The BJPs were isolated and purified and were shown by electrophoretic, gel filtration, and ultracentrifugal analyses to exist as tetramers and dimers. The components in two cases were kappa type and in one lambda type. The kappa tetramers consisted of two covalent dimers and the lambda tetramer two noncovalently bound dimers present in both serum and urine.