Characterization of amyloidogenic immunoglobulin light chains directly from serum by on‐line immunoafﬁnity isolation

Bergen, H. Robert; Abraham, Roshini S.; Johnson, Kenneth L.; Bradwell, Arthur R.; Naylor, Stephen

doi:10.1002/bmc.323

Cited by 19 publications

(11 citation statements)

References 24 publications

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“…These results suggest that the stability of folded light chains needs to be compromised for amyloid formation. Although analysis of LC from serum by immunoaffinity LC-MS/MS (67) did not resolve to what degree disulfide-linked dimers are present in the serum of patients, our data suggest that a reducing milieu may initiate amyloid formation in vivo. Specifically, aggregation was initiated by the reduction of interchain and intermolecular disulfide bonds.…”

Section: Discussioncontrasting

confidence: 71%

Aggregation of Full-length Immunoglobulin Light Chains from Systemic Light Chain Amyloidosis (AL) Patients Is Remodeled by Epigallocatechin-3-gallate

Andrich

Hegenbart

Kimmich

et al. 2017

Journal of Biological Chemistry

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Edited by Paul E. FraserIntervention into amyloid deposition with anti-amyloid agents like the polyphenol epigallocatechin-3-gallate (EGCG) is emerging as an experimental secondary treatment strategy in systemic light chain amyloidosis (AL). In both AL and multiple myeloma (MM), soluble immunoglobulin light chains (LC) are produced by clonal plasma cells, but only in AL do they form amyloid deposits in vivo. We investigated the amyloid formation of patient-derived LC and their susceptibility to EGCG in vitro to probe commonalities and systematic differences in their assembly mechanisms. We isolated nine LC from the urine of AL and MM patients. We quantified their thermodynamic stabilities and monitored their aggregation under physiological conditions by thioflavin T fluorescence, light scattering, SDS stability, and atomic force microscopy. LC from all patients formed amyloid-like aggregates, albeit with individually different kinetics. LC existed as dimers, ϳ50% of which were linked by disulfide bridges. Our results suggest that cleavage into LC monomers is required for efficient amyloid formation. The kinetics of AL LC displayed a transition point in concentration dependence, which MM LC lacked. The lack of concentration dependence of MM LC aggregation kinetics suggests that conformational change of the light chain is rate-limiting for these proteins. Aggregation kinetics displayed two distinct phases, which corresponded to the formation of oligomers and amyloid fibrils, respectively. EGCG specifically inhibited the second aggregation phase and induced the formation of SDS-stable, non-amyloid LC aggregates. Our data suggest that EGCG intervention does not depend on the individual LC sequence and is similar to the mechanism observed for amyloid-␤ and ␣-synuclein. Systemic light chain amyloidosis (AL)2 is the most common form of systemic amyloidosis (1), but it is still a rare disease with an incidence of 6 -7 in 1,000,000 people (2). Underlying AL pathology is a clonal plasma cell disorder, which produces an immunoglobulin light chain in the bone marrow with a unique sequence. These light chains are released into the blood (monoclonal gammopathy). When elevated LC levels in serum overwhelm renal absorption, LC is also found in the urine. Fulllength LC can be isolated from the urine of these patients (3-5).Clonal plasma cell disorders can present different types of pathologies; light chains form amyloid deposits in AL patients. Here, LC fibrils adopt a conformation characterized by highly stable, intramolecular cross-␤-sheets that stain with Congo red (6) and thioflavin T (ThT) (7-9).In contrast, amyloid deposits are not observed in patients suffering from multiple myeloma (MM), a malignant disease characterized by bone marrow failure and bone destruction. The two contrasting pathologies of LC gammopathies raise the question of which factors determine amyloid formation. Amyloid formation could be initiated (a) by the sequences of the individual light chains, (b) by high levels of light chain expression, (c) by p...

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Section: Discussioncontrasting

confidence: 71%

Aggregation of Full-length Immunoglobulin Light Chains from Systemic Light Chain Amyloidosis (AL) Patients Is Remodeled by Epigallocatechin-3-gallate

Andrich

Hegenbart

Kimmich

et al. 2017

Journal of Biological Chemistry

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“…The primary sequence and post-translational modifications of circulating FLC can be described using mass spectrometry; although still experimental, proteomic analysis of these proteins is a promising tool to detect features related to their pathogenicity [98,99].…”

Section: Measurement Of the Amyloidogenic Precursormentioning

confidence: 99%

Biochemical markers in early diagnosis and management of systemic amyloidoses

Lavatelli

Albertini

Fonzo

et al. 2014

Clinical Chemistry and Laboratory Medicine (CCLM)

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Systemic amyloid diseases are characterized by widespread protein deposition as amyloid fibrils. Precise diagnostic framing is the prerequisite for a correct management of patients. This complex process is achieved through a series of steps, which include detection of the tissue amyloid deposits, identification of the amyloid type, demonstration of the amyloidogenic precursor, and evaluation of organ dysfunction/damage. Laboratory medicine plays a central role in the diagnosis and management of systemic amyloidoses, through the quantification of the amyloidogenic precursor and evaluation of end-organ damage using biomarkers.

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“…The methods developed were translated to clinical assays [28, 45], however, these methods require antibody of high specificity, and some mutations may destroy the antigenic epitope leading to reduction or loss of antibody binding. In addition, amyloid proteins may preferentially deposit in tissue leading to low serum levels.…”

Section: Proteomic Investigations In Amyloidosis Typingmentioning

confidence: 99%

Proteomics in Molecular Diagnosis: Typing of Amyloidosis

Loo

Mollee

Renaut

et al. 2011

BioMed Research International

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Amyloidosis is a group of disorders caused by deposition of misfolded proteins as aggregates in the extracellular tissues of the body, leading to impairment of organ function. Correct identification of the causal amyloid protein is absolutely crucial for clinical management in order to avoid misdiagnosis and inappropriate, potentially harmful treatment, to assess prognosis and to offer genetic counselling if relevant. Current diagnostic methods, including antibody-based amyloid typing, have limited ability to detect the full range of amyloid forming proteins. Recent investigations into proteomic identification of amyloid protein have shown promise. This paper will review the current state of the art in proteomic analysis of amyloidosis, discuss the suitability of techniques based on the properties of amyloidosis, and further suggest potential areas of development. Establishment of mass spectrometry aided amyloid typing procedures in the pathology laboratory will allow accurate amyloidosis diagnosis in a timely manner and greatly facilitate clinical management of the disease.

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Characterization of amyloidogenic immunoglobulin light chains directly from serum by on‐line immunoafﬁnity isolation

Cited by 19 publications

References 24 publications

Aggregation of Full-length Immunoglobulin Light Chains from Systemic Light Chain Amyloidosis (AL) Patients Is Remodeled by Epigallocatechin-3-gallate

Aggregation of Full-length Immunoglobulin Light Chains from Systemic Light Chain Amyloidosis (AL) Patients Is Remodeled by Epigallocatechin-3-gallate

Biochemical markers in early diagnosis and management of systemic amyloidoses

Proteomics in Molecular Diagnosis: Typing of Amyloidosis

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