Crystal structure of the C-terminal domain of tubulin-binding cofactor C from Leishmania major

Barrack, K.L.; Fyfe, Paul K.; Hunter, William N.

doi:10.1016/j.molbiopara.2015.05.003

Cited by 5 publications

(2 citation statements)

References 31 publications

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“…Tubulins are members of a family of globular proteins responsible for the cell cytoskeleton [22]. In Leishmania, they interact forming microtubules, which play an important role not only in cell structure and division, but also in keeping the shape and motility of the parasites [23]. They represent housekeeping proteins found in the cytoplasm and nucleus of parasites, being abundantly expressed in both promastigote and amastigote forms, and considered as important components of leishmanial flagella [24].…”

Section: Discussionmentioning

confidence: 99%

Leishmania infantum β-Tubulin Identified by Reverse Engineering Technology through Phage Display Applied as Theranostic Marker for Human Visceral Leishmaniasis

Costa

Alves

Carneiro

et al. 2019

IJMS

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Two Leishmania infantum mimotopes (B10 and C01) identified by phage display showed to be antigenic and immunogenic for visceral (VL) and tegumentary (TL) leishmaniasis; however, their biological targets in the parasites have not been identified. The aim of the present study was to investigate the native antigens expressing both mimotopes, and to use them in distinct immunological assays. For this, a subtractive phage display technology was used, where a combinatorial library of single-chain variable fragments (scFv) was employed and the most reactive monoclonal antibodies for each target were captured, being the target antigens identified by mass spectrometry. Results in immunoblotting and immunoprecipitation assays showed that both monoclonal scFvs antibodies identified the β-tubulin protein as the target antigen in L. infantum. To validate these findings, the recombinant protein was cloned, purified and tested for the serodiagnosis of human leishmaniasis, and its immunogenicity was evaluated in PBMC derived from healthy subjects and treated or untreated VL patients. Results showed high diagnostic efficacy, as well as the development of a specific Th1 immune response in the cell cultures, since higher IFN-γ and lower IL-10 production was found.

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Section: Discussionmentioning

confidence: 99%

Leishmania infantum β-Tubulin Identified by Reverse Engineering Technology through Phage Display Applied as Theranostic Marker for Human Visceral Leishmaniasis

Costa

Alves

Carneiro

et al. 2019

IJMS

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“…The right-handed β-helical CARP domain is also present in several other proteins including tubulin cofactor C (TBCC) and X-linked retinitis pigmentosa 2 (RP2) [24]. However, either TBCC [38] or RP2 [39] lacks a dimerization motif and binds to a ligand as a monomer. Therefore, CAP has adopted a unique dimerization motif to bind to actin monomers.…”

Section: Discussionmentioning

confidence: 99%

The C-terminal dimerization motif of cyclase-associated protein is essential for actin monomer regulation

Iwase

Ono

2016

Biochemical Journal

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Cyclase-associated protein (CAP) is a conserved actin-regulatory protein that functions together with actin-depolymerizing factor (ADF)/cofilin to enhance actin filament dynamics. CAP has multiple functional domains, and the function to regulate actin monomers is carried out by its C-terminal half containing a Wiskott-Aldrich Syndrome protein homology 2 (WH2) domain, a CAP and X-linked retinitis pigmentosa 2 (CARP) domain, and a dimerization motif. WH2 and CARP are implicated in binding to actin monomers and important for enhancing filament turnover. However, a role of the dimerization motif is unknown. Here, we investigated function of the dimerization motif of CAS-2, a CAP isoform in the nematode Caenorhabditis elegans, in actin monomer regulation. CAS-2 promotes ATP-dependent recycling of ADF/cofilin-bound actin monomers for polymerization by enhancing exchange of actin-bound nucleotides. A C-terminal half of CAS-2 (CAS-2C) has nearly as strong activity as full-length CAS-2. Maltose-binding protein (MBP)-tagged CAS-2C is a dimer. However, MBP-CAS-2C with a truncation of either one or two C-terminal β-strands is monomeric. Truncations of the dimerization motif in MBP-CAS-2C nearly completely abolish its activity to sequester actin monomers from polymerization and enhance nucleotide exchange on actin monomers. As a result, these CAS-2C variants, also in the context of full-length CAS-2, fail to compete with ADF/cofilin to release actin monomers for polymerization. CAS-2C variants lacking the dimerization motif exhibit enhanced binding to actin filaments, which is mediated by WH2. Together, these results suggest that the evolutionarily conserved dimerization motif of CAP is essential for its C-terminal region to exert the actin-monomer-specific regulatory function.

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Tubulin Folding and Degradation

Serna

Zabala

2016

Encyclopedia of Life Sciences

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Synthesis of polypeptides inside a cell is an astonishing, complex and very efficient process. To reach the active state, proteins will have to face many different problems that in part are solved by molecular chaperones. Tubulins belong to one of the largest superfamilies of proteins with six conserved members. Different members have different roles although they share a general fold. Tubulins reach their final structure in an intricate pathway where different molecular chaperones are involved. Prefoldin and cytosolic chaperonin containing TCP1/TriC are required in the first part of the pathway, while tubulin folding cofactors (TBCs) are involved in the formation of the α‐ and β‐tubulin heterodimers. Later, their role in tubulin proteostasis as well as their implications in different neurodevelopmental disorders has been documented. Key Concepts Tubulins represent one of the largest families of genes and proteins. α‐ and β‐Tubulin heterodimers constitute the structural and functional subunit of microtubules (MTs). Folding of tubulins follows a sophisticated pathway involving several molecular chaperones. Formation of the αβ‐tubulin heterodimer is an extremely regulated process for it is involved in different lateral and longitudinal as well as microtubule associated proteins (MAPs) interactions. TBCs are molecular chaperones specifically required for αβ‐tubulin dimer formation. TBCs are shown to be involved in the release of tubulin monomers from the chaperonin during the folding pathway. The dissociation constant for the tubulin heterodimer has shown to be 10 −11 M and TBCs are required for an efficient tubulin dimer dissociation. TBCs are involved in tubulin proteostasis through dimer dissociation and tagging for proteasome recognition. The first 3D structure of a complex formed between α‐tubulin and two chaperones involved in the tubulin turnover cycle has been established. Genetic defects on TBCs are responsible of neurodevelopmental diseases.

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Crystal structure of the C-terminal domain of tubulin-binding cofactor C from Leishmania major

Cited by 5 publications

References 31 publications

Leishmania infantum β-Tubulin Identified by Reverse Engineering Technology through Phage Display Applied as Theranostic Marker for Human Visceral Leishmaniasis

Leishmania infantum β-Tubulin Identified by Reverse Engineering Technology through Phage Display Applied as Theranostic Marker for Human Visceral Leishmaniasis

The C-terminal dimerization motif of cyclase-associated protein is essential for actin monomer regulation

Tubulin Folding and Degradation

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