Untitled

̈ger, Hubert A. Ja; Kracker, Bjo ̈rn; Rapp, Lutz

doi:10.1023/a:1026570630964

Cited by 3 publications

(3 citation statements)

References 5 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In the folding intermediate of this single-chain Fv fragment, the inner β-sheet of the V L domain is protected from exchange, while the V H domain has no extended protected nucleus. In a more recent paper, it was shown that the V L domain assists in folding of the V H domain through the formation of a transient unproductive V H -V L domain (52). More details on the folding of llama V HH 's are required to deepen our insight into the structural aspects of the formation of these isolated domains.…”

Section: Discussionmentioning

confidence: 99%

Thermal Unfolding of a Llama Antibody Fragment: A Two-State Reversible Process

et al. 2000

View full text Add to dashboard Cite

Camelids produce functional "heavy chain" antibodies which are devoid of light chains and CH1 domains [Hamers-Casterman, C., et al. (1993) Nature 363, 446-448]. It has been shown that the variable domains of these heavy chain antibodies (the V(HH) fragments) are functional at or after exposure to high temperatures, in contrast to conventional antibodies [Linden van der, R. H. J., et al. (1999) Biochim. Biophys. Acta 1431, 37-44]. For a detailed understanding of the higher thermostability of these V(HH) fragments, knowledge of their structure and conformational dynamics is required. As a first step toward this goal, we report here the essentially complete (1)H and (15)N NMR backbone resonance assignments of a llama V(HH) antibody fragment, and an extensive analysis of the structure at higher temperatures. The H-D exchange NMR data at 300 K indicate that the framework of the llama V(HH) fragment is highly protected with a DeltaG(ex) of >5.4 kcal/mol, while more flexibility is observed for surface residues, particularly in the loops and the two outer strands (residues 4-7, 10-13, and 58-60) of the beta-sheet. The CD data indicate a reversible, two-state unfolding mechanism with a melting transition at 333 K and a DeltaH(m) of 56 kcal/mol. H-D exchange studies using NMR and ESI-MS show that below 313 K exchange occurs through local unfolding events whereas above 333 K exchange mainly occurs through global unfolding. The lack of a stable core at high temperatures, observed for V(HH) fragments, has also been observed for conventional antibody fragments. The main distinction between the llama V(HH) fragment and conventional antibody fragments is the reversibility of the thermal unfolding process, explaining its retained functionality after exposure to high temperatures.

show abstract

Section: Discussionmentioning

confidence: 99%

Thermal Unfolding of a Llama Antibody Fragment: A Two-State Reversible Process

et al. 2000

View full text Add to dashboard Cite

show abstract

“…Previously, we demonstrated that the isolated VL domain of the F11 antibody is functional and does reach complete folding, in contrast to the functional VH-linker-VL fusion protein (scFv fragment F11) that comprises the two domains trapped in a partially unfolded state [22,26]. Plu « ckthun and co-workers provided convincing evidence that premature interactions between partially structured VH and VL domains within an scFv fragment might result in a kinetic trap along the folding pathway [44,45]. Given these data, a partially unfolded conformation for both VL and VH domains within the scFv fragment F11 under physiological conditions can be attributed to premature interactions that the partially structured VH domain establishes with the partner VL domain before the latter attains complete folding.…”

Section: Discussionmentioning

confidence: 99%

Partially structured state of the functional VH domain of the mouse anti‐ferritin antibody F11

et al. 2002

View full text Add to dashboard Cite

An antibody combining site generally involves the two variable domains, VH from the heavy and VL from the light chain. We expressed the individual VH domain of the mouse antihuman ferritin monoclonal antibody F11. The loss of affinity was not dramatic (K a = 4.0U U10 7 M 31 versus 8.6U U10 8 M 31 for the parent antibody) and comparable to that previously observed for other VHs. However, the functional VH domain adopted a partially structured state with a significant amount of distorted secondary and compact yet greatly destabilized tertiary structures, as demonstrated by spectroscopic and calorimetric probes. These data provide the first description for a functional antibody domain that meets all the criteria of a partially structured state. ß 2002 Published by Elsevier Science B.V. on behalf of the Federation of European Biochemical Societies.

show abstract

“…As far as linguistic classifications are concerned, the internal relations of the TG branch of the Tupı ´an family have received much scholarly attention, with different approaches employed to establish them from linguistic data alone. Phonological criteria have been put to use alongside grammatical properties and lexical cognacy, both in "traditional" [7][8][9][10][11][12] and "quantitative" approaches [13][14][15][16][17]. Although these studies agree to a large extent on the topology of the shallower splits, there are still irreconcilable differences in terms of the deepest ones, and much disagreement about their dating.…”

Section: Introductionmentioning

confidence: 99%

Lexical phylogenetics of the Tupí-Guaraní family: Language, archaeology, and the problem of chronology

et al. 2023

View full text Add to dashboard Cite

Tupí-Guaraní is one of the largest branches of the Tupían language family, but despite its relevance there is no consensus about its origins in terms of age, homeland, and expansion. Linguistic classifications vary significantly, with archaeological studies suggesting incompatible date ranges while ethnographic literature confirms the close similarities as a result of continuous inter-family contact. To investigate this issue, we use a linguistic database of cognate data, employing Bayesian phylogenetic methods to infer a dated tree and to build a phylogeographic expansion model. Results suggest that the branch originated around 2500 BP in the area of the upper course of the Tapajós-Xingu basins, with a split between Southern and Northern varieties beginning around 1750 BP. We analyse the difficulties in reconciling archaeological and linguistic data for this group, stressing the importance of developing an interdisciplinary unified model that incorporates evidence from both disciplines.

show abstract

Untitled

Cited by 3 publications

References 5 publications

Thermal Unfolding of a Llama Antibody Fragment: A Two-State Reversible Process

Thermal Unfolding of a Llama Antibody Fragment: A Two-State Reversible Process

Partially structured state of the functional VH domain of the mouse anti‐ferritin antibody F11

Lexical phylogenetics of the Tupí-Guaraní family: Language, archaeology, and the problem of chronology

Contact Info

Product

Resources

About