Effects of dehydroalanine on peptide conformations

Palmer, Darryl Erik; Pattaroni, Christian; Nunami, Ken-ichi; Chadha, Raj K.; Goodman, Murray; Wakamiya, Tateaki; Fukase, Koichi; Horimoto, Shingo; Kitazawa, Manabu

doi:10.1021/ja00040a024

Cited by 109 publications

(80 citation statements)

References 1 publication

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“…Their presence may impose (certain conformational restrictions on this part of the molecule. It has been pointed out that qp-dehydro residues at positions I probably induce the formation of an inverse y-turn centered at position i-1 and stabilized by a C = O ... HN hydrogen bond between residues i -2 and i (Gupta and Chauhan, 1990;Palmer et al, 1992). With respect to its secondary and tertiary structure, it seems likely that epilancin K7 like the other type-A lantibiotics adopts an extended, flexible structure in water (Freund et al, 1991a;van de Ven et al, 1991 ;Lian et al, 1992;Chan et al, 1992).…”

Section: Sequence Determination ; the 'Hi3c-hmbc Approachmentioning

confidence: 99%

Sequence Analysis by NMR Spectroscopy of the Peptide Lantibiotic Epilancin K7 from Staphylococcus epidermidis K7

Kamp

Horstink

Hooven

et al. 1995

European Journal of Biochemistry

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The amino acid sequence of the novel lantibiotic epilancin K7 from Staphylococcus epidermidis K7 was determined by NMR spectroscopy. NMR spectroscopy was used because sequencing by conventional Edman degradation techniques was prohibited by internal sequence blocks owing to the presence of modified residues. Epilancin K7 consists of 31 residues, including two a,P-didehydroalanine (one-letter code U) and two a$-didehydrobutyrine (0) residues, one lanthionine (A-S-A), two p-methyllanthionines (A*-S-A), and six lysines. Epilancin K7 has a molecular mass of 3032 5 1.5 Da. The amino acid sequence of epilancin K7 was derived from both through-space dipolar proton-proton interactions and throughbond scalar proton-carbon interactions as detected by two-dimensional 'H-NOESY, 'H-ROESY and threedimensional 'H-TOCSY-NOESY, and by two-dimensional 'H,'T-heteronuclear multiple-bond correlation spectroscopy, respectively. The sequence is as follows : r s i rshs7 XAUVLKOUIKVAKKYAKGVA*LA* AGANIOGGK.The N-terminal residue X partly resembles an alanine but its exact nature is unclear. The organization of the sulfide-bridge-containing (P-methyl-)lanthionines was revealed by 'H-NMR and 'H,I3C-NMR spectroscopy. Epilancin K7 has a linear structure and a high positive net charge, and therefore is classified as a type-A lantibiotic. NMR analysis of a degraded though still active form of epilancin K7 showed that two N-terminal residues of epilancin K7 were missing, owing to decomposition at the a$-didehydro alanine at position 3 ; it was called the epilancin K7-(3-31)-peptide (peptide fragment of epilancin K7 consisting of positions 3 -31). The usefulness of three-dimensional 'H-TOCSY-NOESY, and two-dimensional 'H,'3C-heter~n~~lear multiple-bond correlation spectroscopy at natural abundance for the study of (modified) polypeptides is demonstrated.Keywords. Bacteriocin; HMBC spectroscopy; lanthionine; post-translational modification; primary structure.Lantibiotics are defined as gene-encoded, ribosomally synthesized antimicrobial polypeptides (19-34 residues, 1800-3500 Da) that contain unusual (P-methyl-)lanthionines and a$-didehydro residues as the result of post-translational modifications Jung and Sahl, 1991;Bierbaum and Sahl, 1993;Hansen, 1993 Abbreviations. A* or Abu,, 2-aminobutyrine or P-methylalanine moiety of /hnethyllanthionine ; Ala,, alanine moiety of lanthionine and P-methyllanthionine ; C', main-chain carbonyl (C =0) carbon ; COSY, correlated spectroscopy; 6, chemical shift; U or Dha, a,b-didehydroalanine ; 0 or Dhb, a$-didehydrobutyrine ; DQ-COSY, double-quantum

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Section: Sequence Determination ; the 'Hi3c-hmbc Approachmentioning

confidence: 99%

Sequence Analysis by NMR Spectroscopy of the Peptide Lantibiotic Epilancin K7 from Staphylococcus epidermidis K7

Kamp

Horstink

Hooven

et al. 1995

European Journal of Biochemistry

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“…Dehydrated residues can enhance or modulate the biological activity of the peptide. For instance, they can play an essential role in peptide activities (35), they can act as inhibitors of biological activities (31,32), or they may alter and improve signal transmission by interaction with receptor or acceptor molecules (29). Furthermore, dehydrated residues can be starting points for further modifications.…”

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confidence: 99%

Sec-Mediated Transport of Posttranslationally Dehydrated Peptides in Lactococcus lactis

Kuipers¹,

Wierenga²,

Rink³

et al. 2006

Appl Environ Microbiol

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Nisin is a lanthionine-containing antimicrobial peptide produced by Lactococcus lactis. Its (methyl)lanthionines are introduced by two posttranslational enzymatic steps involving the dehydratase NisB, which dehydrates serine and threonine residues, and the cyclase NisC, which couples these dehydrated residues to cysteines, yielding thioether-bridged amino acids called lanthionines. The prenisin is subsequently exported by the ABC transporter NisT and extracellularly processed by the peptidase NisP. L. lactis expressing the nisBTC genes can modify and secrete a wide range of nonlantibiotic peptides. Here we demonstrate that in the absence of NisT and NisC, the Sec pathway of L. lactis can be exploited for the secretion of dehydrated variants of therapeutic peptides. Furthermore, posttranslational modifications by NisB and NisC still occur even when the nisin leader is preceded by a Sec signal peptide or a Tat signal peptide 27 or 44 amino acids long, respectively. However, transport of fully modified prenisin via the Sec pathway is impaired. The extent of NisB-mediated dehydration could be improved by raising the intracellular concentration NisB or by modulating the export efficiency through altering the signal sequence. These data demonstrate that besides the traditional lantibiotic transporter NisT, the Sec pathway with an established broad substrate range can be utilized for the improved export of lantibiotic enzyme-modified (poly)peptides.

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“…Furthermore, it is known that the group can interact through hyperconjugation with a carbonyl group, in a manner akin to a vinyl group. 7 The N-C  -C(=O) bond angle of the ACCa is usually close to 116 o , 6 which is closer to the theoretical 120 o of the sp 2 hybridised system of the Dha than the tetrahedral bond angle of 109.5 o (Figure 2). The proposed Dha5 replacements will ultimately give a greater understanding of how and why structural changes in that region might impact the overall biological activity.…”

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confidence: 51%

“…2 However, its stability and solubility decrease considerably at physiological pH which precludes its use as a human therapeutic. The A and B rings of the N-terminus of nisin (nisin [3][4][5][6][7][8][9][10][11][12] ) are critical to its overall biological activity where it binds to the lipid II moiety inhibiting the biosynthesis of the bacterial cell wall. 3 It is known that at physiological pH the two Dha residues at positions 5 and 33 are cleaved by hydrolysis.…”

mentioning

confidence: 99%

“…NMR studies by the Murray group showed that the overall conformation of the A-ring of nisin changed substantially when the Dha5 residue was replaced by either a L-Ala or D-Ala moiety. 6 For our studies one of the crucial aspects of the endeavour would be the synthesis of useful A-ring analogues of nisin, where Dha5 has been replaced, to study how changes affect both the conformations adopted and the resulting stability of the peptides. Herein, the initial synthetic targets, where the replacement of Dha5 with glycine, L-alanine, L-serine and L-phenylalanine residues is described.…”

mentioning

confidence: 99%

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Solid-phase peptide synthesis of analogues of the N-terminus A-ring fragment of the lantibiotic nisin: Replacements for the dehydroalanine (Dha) residue at position 5 and the first incorporation of a thioamide residue

Manzor¹,

Proinsias

Kelleher³

2017

Tetrahedron Letters

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Effects of dehydroalanine on peptide conformations

Cited by 109 publications

References 1 publication

Sequence Analysis by NMR Spectroscopy of the Peptide Lantibiotic Epilancin K7 from Staphylococcus epidermidis K7

Sequence Analysis by NMR Spectroscopy of the Peptide Lantibiotic Epilancin K7 from Staphylococcus epidermidis K7

Sec-Mediated Transport of Posttranslationally Dehydrated Peptides in Lactococcus lactis

Solid-phase peptide synthesis of analogues of the N-terminus A-ring fragment of the lantibiotic nisin: Replacements for the dehydroalanine (Dha) residue at position 5 and the first incorporation of a thioamide residue

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