Fifty years of coiled-coils and α-helical bundles: A close relationship between sequence and structure

Parry, David; Fraser, R.D.B.; Squire, John M.

doi:10.1016/j.jsb.2008.01.016

Cited by 246 publications

(223 citation statements)

References 89 publications

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“…Many proteins assemble or interact with their partners through coiled-coil domains, which can associate homophilically and/or heterophilically to form dimers, trimers, or tetramers (26)(27)(28). Our work shows that TRPP2 forms a homotrimer through a coiled-coil domain (amino acids F839-A873) in the C terminus.…”

Section: Discussionmentioning

confidence: 77%

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Structural and molecular basis of the assembly of the TRPP2/PKD1 complex

Ulbrich

et al. 2009

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

162

120

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Mutations in PKD1 and TRPP2 account for nearly all cases of autosomal dominant polycystic kidney disease (ADPKD). These 2 proteins form a receptor/ion channel complex on the cell surface. Using a combination of biochemistry, crystallography, and a single-molecule method to determine the subunit composition of proteins in the plasma membrane of live cells, we find that this complex contains 3 TRPP2 and 1 PKD1. A newly identified coiled-coil domain in the C terminus of TRPP2 is critical for the formation of this complex. This coiled-coil domain forms a homotrimer, in both solution and crystal structure, and binds to a single coiled-coil domain in the C terminus of PKD1. Mutations that disrupt the TRPP2 coiled-coil domain trimer abolish the assembly of both the full-length TRPP2 trimer and the TRPP2/PKD1 complex and diminish the surface expression of both proteins. These results have significant implications for the assembly, regulation, and function of the TRPP2/PKD1 complex and the pathogenic mechanism of some ADPKD-producing mutations.autosomal dominant polycystic kidney disease ͉ single-molecule imaging ͉ stoichiometry ͉ transient receptor potential channel ͉ X-ray crystallography

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Section: Discussionmentioning

confidence: 77%

“…3 C and F). This coiled-coil has many of the hallmarks of a canonical coiled-coil (26)(27)(28), in particular, the characteristic heptad repeat with hydrophobic residues at the first (i.e., a) and fourth (i.e., d) positions, and with charged residues at the fifth (i.e., e) and seventh (i.e., g) positions (Fig. 3B).…”

Section: Discussionmentioning

confidence: 99%

Structural and molecular basis of the assembly of the TRPP2/PKD1 complex

Ulbrich

et al. 2009

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

162

120

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“…Early work on fibrous α-keratin predicted formation of extended structures with coiled-coil α-helices, which was later witnessed in a variety of proteins (25). Similarly, a model for fibrous collagen with three left-handed polyproline type II (PPII) helices forming a right-handed supercoil was proposed (26) and then observed structurally (27).…”

Section: Discussionmentioning

confidence: 96%

“…Finally, the carboxy terminus contains the LPxTG sortase motif for covalent anchorage to the cell wall (13). S. mutans AgI/II possesses both low-and high-affinity binding sites for salivary agglutinin (SAG) (14), a 600-kDa oligomeric protein complex containing glycoprotein 340 (440 kDa), sIgA (25,59, and 88 kDa), and an unknown 80-kDa polypeptide (15). Studies aimed at developing active and passive immune therapies have analyzed segments of Ag I/II that contribute to bacterial adherence and cariogenicity (2,16).…”

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confidence: 99%

Elongated fibrillar structure of a streptococcal adhesin assembled by the high-affinity association of α- and PPII-helices

Larson

Rajashankar

Patel

et al. 2010

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

175

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Streptococcus mutans antigen I/II (AgI/II) is a cell surface-localized protein adhesin that interacts with salivary components within the salivary pellicle. AgI/II contributes to virulence and has been studied as an immunological and structural target, but a fundamental understanding of its underlying architecture has been lacking. Here we report a high-resolution (1.8 Å) crystal structure of the A 3 VP 1 fragment of S. mutans AgI/II that demonstrates a unique fibrillar form (155 Å) through the interaction of two noncontiguous regions in the primary sequence. The A 3 repeat of the alanine-rich domain adopts an extended α-helix that intertwines with the P 1 repeat polyproline type II (PPII) helix to form a highly extended stalk-like structure heretofore unseen in prokaryotic or eukaryotic protein structures. Velocity sedimentation studies indicate that fulllength AgI/II that contains three A/P repeats extends over 50 nanometers in length. Isothermal titration calorimetry revealed that the high-affinity association between the A 3 and P 1 helices is enthalpically driven. Two distinct binding sites on AgI/II to the host receptor salivary agglutinin (SAG) were identified by surface plasmon resonance (SPR). The current crystal structure reveals that AgI/II family proteins are extended fibrillar structures with the number of alanine-and proline-rich repeats determining their length.bacterial adhesion | dental caries | Streptococcus | x-ray crystallography | fibrous proteins S treptococcus mutans is the causative agent of human dental caries (1) and its protein adhesin antigen I/II (AgI/II) is a known target of protective immunity (2). AgI/II family molecules are expressed by numerous oral streptococci (3) and homologs have also been identified in the invasive pathogens Streptococcus pyogenes and Streptococcus agalactiae (4) (Fig. S1). In addition to mediating adhesion to the tooth surface (5), AgI/II influences biofilm formation (6), promotes collagen-dependent bacterial invasion of dentin (7), and mediates adherence to human epithelial cells (8). Elimination of AgI/II results in decreased virulence (9), but despite three decades of study, a mechanistic understanding of the functional properties of the molecule has been stymied by a lack of understanding of its structure.Originally identified as AgI/II (10) (also called P1, PAc, or SpaP), members of this protein family contain between 1310 and 1653 amino acids (aa) beginning with an amino-terminal signal motif that directs secretion, followed by the A, V, and P regions (Fig. 1A). The A region typically consists of 3-4 alanine-rich repeats (82 residues each) with 23-30% alanine content. The P region has 3-4 proline-rich repeats (39 residues each) with ∼35% proline content. Nested between the A and P repeats is a segment commonly referred to as the V or variable region, which contains within it a stretch of ∼100 amino acids where most of the sequence variation among S. mutans AgI/II molecules is clustered (11). The crystal structure of the V region adopts a globular β-s...

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“…8 The regularity and simplicity of the motif has proved to be a very good candidate for de novo protein design in Abbreviations: ECFP, enhanced cyan fluorescent protein; FRET, fluorescence resonance energy transfer; TEV, tobacco etch virus. the past; 2,9-15 for example, properties like the association constant, specificity, and oligomerization state of interacting peptides in solution could be controlled to some extent.…”

Section: Introductionmentioning

confidence: 99%

Induced heterodimerization and purification of two target proteins by a synthetic coiled‐coil tag

Fernández-Rodríguez

Marlovits

2012

Protein Science

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A synthetic de novo designed heterodimeric coiled-coil was used to copurify two target fluorescent proteins, Venus and enhanced cyan fluorescent protein (ECFP). The coiled-coil consists of two 21-amino acid repetitive sequences, (EIAALEK) 3 and (KIAALKE) 3 , named E3 and K3, respectively. These sequences were fused to the C-termini of ECFP or Venus followed by either a strep-or a his-tag, respectively, for affinity purification. Mixed lysates of Venus-K3 and ECFP-E3 were subjected to consecutive affinity purification and showed highly specific association between the coiled-coil pair by SDS-PAGE, gel filtration, isothermal titration calorimetry (ITC), and fluorescence resonance energy transfer (FRET). The tagged proteins eluted as heterodimers at the concentrations tested. FRET analysis further showed that the coiled-coil pair was stable in buffers commonly used for protein purification, including those containing high salt concentration and detergent. This study shows that the E3/K3 pair is very well suited for the copurification of two target proteins expressed in vivo because of its high specificity: it forms exclusively heterodimers in solution, it does not interact with any cellular proteins and it is stable under different buffer conditions.

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Fifty years of coiled-coils and α-helical bundles: A close relationship between sequence and structure

Cited by 246 publications

References 89 publications

Structural and molecular basis of the assembly of the TRPP2/PKD1 complex

Structural and molecular basis of the assembly of the TRPP2/PKD1 complex

Elongated fibrillar structure of a streptococcal adhesin assembled by the high-affinity association of α- and PPII-helices

Induced heterodimerization and purification of two target proteins by a synthetic coiled‐coil tag

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