Conformation dependent monoclonal antibodies distinguish different replicating strains or conformers of prefibrillar Aβ oligomers

Kayed, Rakez; Canto, Isabel; Breydo, Leonid; Rasool, Suhail; Lukácsovich, Tamás; Wu, Jessica; Albay, Ricardo; Pensalfini, Anna; Yeung, Stephen T.; Head, Elizabeth; Marsh, J. Lawrence; Glabe, Charles G.

doi:10.1186/1750-1326-5-57

Cited by 143 publications

(160 citation statements)

References 30 publications

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“…Comparison of LFAOs with Other Oligomers Indicates Similarities and Differences-There are only a handful of reports on the replication property of A␤ oligomers, and of those, the reports by Glabe and co-workers (20,28,37) are the most recent. They identified three specific soluble oligomer subtypes classified mainly on the basis of their immunoreactivity toward conformation-specific antibodies.…”

Section: Discussionmentioning

confidence: 99%

“…They identified three specific soluble oligomer subtypes classified mainly on the basis of their immunoreactivity toward conformation-specific antibodies. These subtypes are PFOs, fibrillar oligomers (FOs), and APFs, and they are recognized by the conformation-specific antibodies A11, OC, and ␣PF, respectively (28). Among these, both PFOs and FOs showed a self-replication property by converting monomers into oligomers of the same type (28,38).…”

Section: Discussionmentioning

confidence: 99%

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Specific Soluble Oligomers of Amyloid-β Peptide Undergo Replication and Form Non-fibrillar Aggregates in Interfacial Environments

Kumar

Paslay

Lyons

et al. 2012

Journal of Biological Chemistry

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Background: Oligomers of amyloid-␤ peptides are implicated in the etiology of Alzheimer disease. Results: Specific "off-pathway" oligomers of A␤42 show unique replication properties upon interacting with monomers. Conclusion:The results indicate that oligomers that are formed along pathways outside the fibril formation pathway may undergo replication. Significance: Mechanistic details of A␤ soluble oligomers will enable better understanding of Alzheimer disease pathology.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Specific Soluble Oligomers of Amyloid-β Peptide Undergo Replication and Form Non-fibrillar Aggregates in Interfacial Environments

Kumar

Paslay

Lyons

et al. 2012

Journal of Biological Chemistry

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“…7B, all CRES subgroup proteins possessed structures characteristic of amyloid including bundles of fibrils, protofibrils with a beads-on-a-string appearance (CRES), large branched polygons (CRES3) and bundles of fibrils/films assembling into higher ordered structures (CRES2, cystatin E2). Recombinant proteins diluted into aqueous buffer were also examined for amyloid by spotting the samples on to nitrocellulose and carrying out dot blot analysis using conformation-dependent antibodies that recognize the immature forms of amyloid (anti-oligomer A11 antibody) and the mature forms of amyloid (anti-fibrillar OC antibody) (Kayed et al, 2010). The antifibrillar OC antibody bound to all CRES subgroup proteins suggesting the presence of amyloid fibrils, while A11 bound only to CRES and cystatin E2 suggesting they also contained earlier, less mature, forms of amyloid including protofibrils and oligomers (Fig.…”

Section: Cres Subgroup Proteins Form Amyloid In Vitro and In Vivomentioning

confidence: 99%

Cystatin-related epididymal spermatogenic subgroup members are part of an amyloid matrix and associated with extracellular vesicles in the mouse epididymal lumen

et al. 2016

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STUDY QUESTION: Do the CRES (cystatin-related epididymal spermatogenic) subgroup members, including CRES2, CRES3 and cystatin E2, contribute to the formation of a nonpathological, functional amyloid matrix in the mouse epididymal lumen?SUMMARY ANSWER: CRES2, CRES3 and cystatin E2 self-assemble with different aggregation properties into amyloids in vitro, are part of a common amyloid matrix in the mouse epididymal lumen and are present in extracellular vesicles.WHAT IS KNOWN ALREADY: Although previously thought only to be pathological, accumulating evidence has established that amyloids, which are highly ordered protein aggregates, can also carry out functional roles in the absence of pathology. We previously demonstrated that nonpathological amyloids are present in the epididymis; specifically, that the reproductive cystatin CRES forms amyloid and is present in the mouse epididymal lumen in a film-like amyloid matrix that is intimately associated with spermatozoa. Because the related proteins CRES2, CRES3 and cystatin E2 are also expressed in the epididymis, the present studies were carried out to determine if these proteins are also amyloidogenic in vitro and in vivo and thus may coordinately function with CRES as an amyloid structure. STUDY DESIGN, SAMPLES/MATERIALS, METHODS:The epididymides from CD1 and Cst8 (CRES)129SvEv/B6 gene knockout (KO) and wild-type mice and antibodies that specifically recognize each CRES subgroup member were used for immunohistochemical and biochemical analyzes of CRES subgroup proteins. Methods classically used to identify amyloid, including the conformation-dependent dyes thioflavin S (ThS) and thioflavin T (ThT), conformation-dependent antibodies, protein aggregation disease ligand (which binds any amyloid independent of sequence) and negative stain electron microscopy (EM) were carried out to examine the amyloidogenic properties of CRES subgroup members. Immunofluorescence analysis and confocal microscopy were used for colocalization studies. MAIN RESULTS AND THE ROLE OF CHANCE:Immunoblot and immunofluorescence analyzes showed that CRES2, CRES3 and cystatin E2 were primarily found in the initial segment and intermediate zone of the epididymis and were profoundly downregulated in epididymides from CRES KO mice, suggesting integrated functions. Except for CRES3, which was only detected in a particulate form, proteins were present in the epididymal lumen in both soluble and particulate forms including in a film-like matrix and in extracellular vesicles. The use of amyloid-specific reagents determined that all CRES subgroup members were present as amyloids and colocalized to a common amyloid matrix present in the epididymal lumen. Negative stain EM, dot blot analysis and ThT plate assays showed that recombinant CRES2, CRES3 and cystatin E2 formed amyloid in vitro, albeit with different aggregation properties. Together, our studies demonstrate that a unique amyloid matrix composed of the CRES family of reproductive-specific cystatins and cystatin C is a normal component of the mouse ...

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“…A breakthrough in this area has been the development of conformation-specific antibodies that selectively recognize uniquely folded conformers of amyloidogenic proteins (3)(4)(5)(6)(7)(8)(9)(10)(11)(12)(13). Indeed, multiple conformation-specific antibodies have been reported that recognize structural features within amyloidogenic oligomers (5) and fibrils (4,6,8) in a sequence-independent manner.…”

mentioning

confidence: 99%

Structure-based design of conformation- and sequence-specific antibodies against amyloid β

Perchiacca

Ladiwala

Bhattacharya

et al. 2011

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

128

134

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Conformation-specific antibodies that recognize aggregated proteins associated with several conformational disorders (e.g., Parkinson and prion diseases) are invaluable for diagnostic and therapeutic applications. However, no systematic strategy exists for generating conformation-specific antibodies that target linear sequence epitopes within misfolded proteins. Here we report a strategy for designing conformation-and sequence-specific antibodies against misfolded proteins that is inspired by the molecular interactions governing protein aggregation. We find that grafting small amyloidogenic peptides (6-10 residues) from the Aβ42 peptide associated with Alzheimer's disease into the complementarity determining regions of a domain (V H ) antibody generates antibody variants that recognize Aβ soluble oligomers and amyloid fibrils with nanomolar affinity. We refer to these antibodies as gammabodies for grafted amyloid-motif antibodies. Gammabodies displaying the central amyloidogenic Aβ motif ( 18 VFFA 21 ) are reactive with Aβ fibrils, whereas those displaying the amyloidogenic C terminus ( 34 LMVGGVVIA 42 ) are reactive with Aβ fibrils and oligomers (and weakly reactive with Aβ monomers). Importantly, we find that the grafted motifs target the corresponding peptide segments within misfolded Aβ conformers. Aβ gammabodies fail to cross-react with other amyloidogenic proteins and scrambling their grafted sequences eliminates antibody reactivity. Finally, gammabodies that recognize Aβ soluble oligomers and fibrils also neutralize the toxicity of each Aβ conformer. We expect that our antibody design strategy is not limited to Aβ and can be used to readily generate gammabodies against other toxic misfolded proteins.misfolding | beta-amyloid | protein engineering A hallmark of protein misfolding disorders is that polypeptides of unrelated sequence fold into similar oligomeric and fibrillar assemblies that are cytotoxic (1). The structures of these enigmatic conformers have captured the imagination of many investigators who have sought to explain the molecular basis of proteotoxicity in conformational disorders such as Alzheimer's disease. Because misfolded proteins are typically refractory to structural methods such as X-ray crystallography and solution NMR, few high-resolution structures of full-length misfolded proteins have been reported (ref. 2 and references therein). The structures of oligomeric intermediates have proven especially difficult to characterize because these conformers are labile, transient, and, in many cases, heterogeneous.Given the complexity of high-resolution structural analysis of misfolded proteins, alternative biochemical approaches are critical for understanding structure-function relationships of aggregated proteins. A breakthrough in this area has been the development of conformation-specific antibodies that selectively recognize uniquely folded conformers of amyloidogenic proteins (3-13). Indeed, multiple conformation-specific antibodies have been reported that recognize structural features ...

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Conformation dependent monoclonal antibodies distinguish different replicating strains or conformers of prefibrillar Aβ oligomers

Cited by 143 publications

References 30 publications

Specific Soluble Oligomers of Amyloid-β Peptide Undergo Replication and Form Non-fibrillar Aggregates in Interfacial Environments

Specific Soluble Oligomers of Amyloid-β Peptide Undergo Replication and Form Non-fibrillar Aggregates in Interfacial Environments

Cystatin-related epididymal spermatogenic subgroup members are part of an amyloid matrix and associated with extracellular vesicles in the mouse epididymal lumen

Structure-based design of conformation- and sequence-specific antibodies against amyloid β

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