Thermally induced conformational transitions of Bence-Jones protein IVA and its proteolytic fragments

Zav’yalov, V.P.; Troitsky, G.V.; Хечинашвили, Н. Н.; Privalov, Peter L.

doi:10.1016/0005-2795(77)90218-5

Cited by 24 publications

(11 citation statements)

References 19 publications

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“…Thermal unfolding of the isolated VL domain of the anti‐ferritin antibody F11 occurred with a T m around 60°C [16], consistent with what was reported for the human VL domain proteolytically derived from the Bence Jones protein IVA [36]. For VL‐barnase at neutral pH, the second peak around 60°C is therefore attributable to the melting of the VL module.…”

Section: Resultssupporting

confidence: 83%

Independent folding and conformational changes of the barnase module in the VL‐barnase immunofusion: calorimetric evidence

2003

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Although stability is critical for in vivo application of immunotoxins, a thermodynamic description of their folding/ stability is still lacking. We applied di¡erential scanning calorimetry (DSC) to RNase-based immunofusion comprising barnase, cytotoxic RNase from Bacillus amyloliquefaciens, fused to the light chain variable domain (VL) of anti-human ferritin antibody F11. By analyzing DSC curves recorded with or without preheating and addition of the barnase-stabilizing ligand guanosine 3P P-monophosphate, we (i) assigned two well-resolved thermal transitions to the VL and barnase modules of VL-barnase, (ii) demonstrated independent folding of these two modules, and (iii) showed altered stability of the barnase module, which resulted from the dimeric state of VL-barnase. ß

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Section: Resultssupporting

confidence: 83%

Independent folding and conformational changes of the barnase module in the VL‐barnase immunofusion: calorimetric evidence

2003

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“…Similar data were also obtained for the LUS protein and its fragments. The data suggest a block structure of the protein because the calorimetric enthalpy is much higher than the effec tive one (calculated also from optical curves of the melt ing, see table) that is specific for the Bence Jones proteins [9,10,18]. For interpretation of the results, it was neces sary to establish just what domains could form the blocks and also to determine thermodynamic parameters of these cooperative structures.…”

Section: Formation In Solution By Variable Domains Of Proteins Tim Anmentioning

confidence: 93%

“…The limited proteolysis procedures leading to cleav age of the peptide bond between the constant and variable domains and the subsequent separation of constant (F b fragment) and variable (F V fragment) parts of the protein were performed in general as described in work [18]. But because the yield of variable domains in the case of pro tein LUS was decreased, the method was modified as fol lows: not the isolated protein LUS was subjected to limit ed proteolysis, but its complex with the Fab fragment [9].…”

Section: Methodsmentioning

confidence: 99%

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Role of cis- and trans-interactions in manifestations of amyloidogenic properties of variable domains of Bence-Jones proteins TIM and LUS

Tishchenko

2013

Biochemistry Moscow

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Intact Bence-Jones proteins TIM and LUS under simulated physiological conditions (10 mM phosphate buffer, pH 7.0, 100 mM NaCl, 37°C) did not display amyloidogenic properties. However, their isolated variable domains exhibit these qualities in full measure. Therefore, both intact proteins and their variable domains were studied using a complex of physical methods (scanning microcalorimetry, analytical centrifugation, optics) that allowed us to assess the stability of their tertiary and quaternary structures. The experimentally obtained thermodynamic functions indicated that the stability of isolated variable domains of TIM and LUS was comparable to the stability of similar domains in amyloidogenic proteins described earlier. However, inside the whole protein their stability was comparable to the stability of VL domains of ordinary Bence-Jones proteins. The decreased stability of the isolated variable domains of TIM and LUS was shown to be due both to weak interactions between a pair of variable domains (trans-interaction) and to a natural lack of interaction with the constant domains (cis-interaction).

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“…It is interesting to note that in most of the studies, the thermal unfolding of Bence-Jones Proteins, free MM LCs, AL LC, and recombinant LCs (encompassing the V L and the C L domain, also called FL) has been described as an apparent 2-state process, showing a single irreversible sigmoidal transition, often followed by protein aggregation [7–11, 14–16]. There is only one report of multiple unfolding transitions for the LC thermal unfolding performed at pH 3.23 [17]. …”

Section: Introductionmentioning

confidence: 99%

Kinetic stability and sequence/structure studies of urine-derived Bence-Jones proteins from multiple myeloma and light chain amyloidosis patients

Blancas‐Mejia

Martin

Williams

et al. 2017

Biophysical Chemistry

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It is now accepted that the ability of a protein to form amyloid fibrils could be associated both kinetic and thermodynamic protein folding parameters. A recent study from our laboratory using recombinant full-length (encompassing the variable and constant domain) immunoglobulin light chains found a strong kinetic control of the protein unfolding for these proteins. In this study, we are extending our analysis by using urine-derived Bence Jones proteins (BJPs) from five patients with light chain (AL) amyloidosis and four patients with Multiple Myeloma (MM). We observed lower stability in κ proteins compared to λ proteins (for both MM and AL proteins) in agreement with previous studies. The kinetic component of protein stability is not a universal feature of BJPs and the hysteresis observed during refolding reactions could be attributed to the inability of the protein to refold all domains. The most stable proteins exhibited 3-state unfolding transitions. While these proteins do not refold reversibly, partial refolding shows 2-state partial refolding transitions, suggesting that one of the domains (possibly the variable domain) does not refold completely. Sequences were aligned with their respective germlines and the location and nature of the mutations were analyzed. The location of the mutations were analyzed and compared with the stability and amyloidogenic properties for the proteins in this study, increasing our understanding of light chain unfolding and amyloidogenic potential.

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Thermally induced conformational transitions of Bence-Jones protein IVA and its proteolytic fragments

Cited by 24 publications

References 19 publications

Independent folding and conformational changes of the barnase module in the VL‐barnase immunofusion: calorimetric evidence

Independent folding and conformational changes of the barnase module in the VL‐barnase immunofusion: calorimetric evidence

Role of cis- and trans-interactions in manifestations of amyloidogenic properties of variable domains of Bence-Jones proteins TIM and LUS

Kinetic stability and sequence/structure studies of urine-derived Bence-Jones proteins from multiple myeloma and light chain amyloidosis patients

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