The Construction of New Proteins. Part III. Artificial folding units by assembly of amphiphilic secondary structures on a template

Mutter, Manfred; Altmann, Eva; Altmann, Karl‐Heinz; Hersperger, René; Koziej, Pjotr; Nebel, K.; Tuchsecherer, Gabriele; Vuilleumier, Stéphane; Gremlich, Hans‐Ulrich; Müller, Klaus

doi:10.1002/hlca.19880710419

Cited by 94 publications

(42 citation statements)

References 38 publications

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“…The templates have included an octapeptide sequence, designed to form a short stretch of antiparallel i-sheet [5], a decapeptide sequence, cyclized by a disulfide bridge between a C-and a N-terminal Cys residue [6], as well as a cyclic decapeptide [7]. Other research groups have applied the TASP concept to Abbreviations: Aoa, aminooxyacetyl; APCI, atmospheric pressure chemical ionization; BAL, backbone amide linker; D-Galp, D-galactopyranose; DIPCDI, N,N%-diisopropylcarbodiimide; DIEA, N,N-diisopropylethylamine; o-PALdehyde, 5-(2-formyl-3,5-dimethoxyphenoxy)valeric acid; TASP, template-assembled synthetic protein; Amino acid symbols denote the L-configuration.…”

Section: Introductionmentioning

confidence: 99%

Carbopeptides: chemoselective ligation of peptide aldehydes to an aminooxy-functionalized D-galactose template

Brask

Jensen

2000

J. Peptide Sci.

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Section: Introductionmentioning

confidence: 99%

Carbopeptides: chemoselective ligation of peptide aldehydes to an aminooxy-functionalized D-galactose template

Brask

Jensen

2000

J. Peptide Sci.

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“…The de novo design of redox proteins is an attractive approach to investigate the factors involved in electron transfer (ET) and to create proteins with novel functions. Synthetic peptides have been linked sequentially (1) or assembled by templates like transition metals (2), porphyrins (3), carbohydrates (4), and peptides (5). In particular, a cyclic peptide template with orthogonally protected amino acids provides selective ligation and a well-defined relative orientation of the peptide elements (6).…”

mentioning

confidence: 99%

Modular synthesis of de novo- designed metalloproteins for light-induced electron transfer

Rau

DeJonge

Haehnel

1998

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

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The design and chemical synthesis of two de novo four-helix bundle proteins is described; each protein has two bound cofactors. Their construction from purified peptides is based on the modular assembly of different amphiphilic helices by chemoselective coupling to a cyclic peptide template. In the hydrophobic interior of the antiparallel four-helix bundle these proteins contain a heme in a binding pocket with two ligating histidine residues. A rutheniumtris(bipyridine) complex is covalently bound to different positions at the hydrophilic side of one of the heme-binding helices. Laser-induced electron transfer across the varied distance through this helix has been studied and compared with a pathway analysis. The UV-visible, CD, and mass spectra are consistent with the structure and orientation predetermined by the template.Redox proteins represent the largest group of enzymes. Their cofactors or redox centers are embedded in structural domains with a wide variety of functions, including light-induced charge separation and electron transport. The size of a domain has come within reach of chemical methods by recent developments in peptide synthesis and chemoselective ligation to link unprotected peptides. The de novo design of redox proteins is an attractive approach to investigate the factors involved in electron transfer (ET) and to create proteins with novel functions. Synthetic peptides have been linked sequentially (1) or assembled by templates like transition metals (2), porphyrins (3), carbohydrates (4), and peptides (5). In particular, a cyclic peptide template with orthogonally protected amino acids provides selective ligation and a well-defined relative orientation of the peptide elements (6). Such a branched design of a template-assembled synthetic protein avoids the folding problem encountered in the association of linear peptides (7) and increases the stability of the folded structure (8). The binding of heme groups to bundles of amphiphilic linear helices has been extensively investigated (9, 10). For studies of the ET, different cofactors have been bound to a single helix (11) or bundles of two (12) and three helices (13). Light-induced ET through metalloproteins like cytochromes and myoglobins (14) has been investigated after modification of surface-exposed amino acid residues by ruthenium complexes. Electron tunneling through a protein (15) is a matter of discussion and has been approximated by a homogeneous barrier to ET in complexes like photosynthetic reaction centers (16). However, different regions of a given protein can indeed display different barriers to ET, as shown by an analysis of pathways through the protein (17). We have extended our model of cytochrome b termed MOP1 with a template-based antiparallel four-helix bundle with two different helices (18) to one with three different helices capable of light-induced ET. One of the two heme-binding amphiphilic helices carries besides the ligating histidine a cysteine at the hydrophilic side to which a ruthenium tris-bipyridine comp...

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“…24 To address these issues, Mutter et al initiated the fusion of template design with de novo peptides to create ''template-assembled synthetic proteins'' (TASPs). 25,26 Appropriately designed TASPs favor helical bundle formation and mitigate the unfavorable entropic cost from bringing four helices together. The template Mutter et al employed was peptidic in nature.…”

Section: Introductionmentioning

confidence: 99%

Four‐helix bundle cavitein reveals middle leucine as linchpin

2007

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A template-assembled de novo four-helix bundle is used to examine the hydrophobic effect within the bundle interior. Leu to Ala variants of the basis sequence GG-EELLKKLEELLKKG were characterized by GuHCl denaturation, NMR dispersion, and N-H/D exchange experiments. The results show that the middle leucine (L7) is imperative in maintaining bundle stability. The average leucine was found to contribute 1.8 kcal mol(-1) toward stability, whereas the middle leucines contribute 2.7 kcal mol(-1) each. Substituting alanine into the middle position (7) constitutes a striking 95% reduction of the overall cavitein stability.

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The Construction of New Proteins. Part III. Artificial folding units by assembly of amphiphilic secondary structures on a template

Cited by 94 publications

References 38 publications

Carbopeptides: chemoselective ligation of peptide aldehydes to an aminooxy-functionalized D-galactose template

Carbopeptides: chemoselective ligation of peptide aldehydes to an aminooxy-functionalized D-galactose template

Modular synthesis of de novo- designed metalloproteins for light-induced electron transfer

Four‐helix bundle cavitein reveals middle leucine as linchpin

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