Assignment of amino acids in peptides by correlation of α-hydrogen and carbonyl carbon-13 resonances

Wynants, C.; Hallenga, Klaas; Binst, G. Van; Michel, Alain; Zanen, J

doi:10.1016/0022-2364(84)90237-3

Cited by 38 publications

(19 citation statements)

References 26 publications

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“…oxygen, something which is not possible with INADEQUATE. In previous applications this experiment has been used to assign the spectra of compounds of established structure (10,11,(17)(18)(19), but the present results clearly demonstrate its potential in the field of structure elucidation of new compounds. It was to test this potential that the present investigation was carried out by the spectroscopic group before any other structural information concerning the compound in hand was available to them.…”

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confidence: 69%

Isolation of 5-methoxy-2,2-dimethyl-1-2H-benzopyran-6-propanoic acid methyl ester and characterization by two-dimensional nuclear magnetic resonance spectroscopy

Jacobs¹,

Ramdayal²,

Reynolds³

et al. 1986

Can. J. Chem.

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confidence: 69%

Isolation of 5-methoxy-2,2-dimethyl-1-2H-benzopyran-6-propanoic acid methyl ester and characterization by two-dimensional nuclear magnetic resonance spectroscopy

Jacobs¹,

Ramdayal²,

Reynolds³

et al. 1986

Can. J. Chem.

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“…Additionally this technique yields a sequence analysis and can be used for peptide sequencing in favorable cases. After completion of this work, the same technique was also reported elsewhere [62]. More recently we developed the 'H,"C-COLOC experiment as a more convenient way to assign CO signals [%I.…”

mentioning

confidence: 86%

Peptide conformations. Part 30. Assignment of the ¹H‐, ¹³C‐, and ¹⁵N‐NMR spectra of cyclosporin A in CDCl₃ and C₆D₆ by a combination of homo‐ and heteronuclear two‐dimensional techniques

Kessler

Loosli

Oschkinat

1985

Helvetica Chimica Acta

159

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The 1H‐, 13C‐, and 15N‐NMR spectra of the immunosuppressive cyclic undecapeptide cyclosporin A (1) have been analyzed at 300 MHz in CDCl3, C6D6, and mixtures of these solvents. A combination of different homo‐ and heteronuclear two‐dimensional NMR techniques enable complete assignment of all H‐, C‐ and 4 N‐signals. Recognition of the proton spin systems has been achieved via 1H,1H–COSY and double‐quantum‐1H‐NMR spectroscopy. NOESY spectra yield some sequence assignments, but two techniques using coupling across amide bonds have been applied to get independent assignments of all amino acids in the sequence: (i) An 1H,1H‐COSY spectrum optimized for small coupling constants enables the detection of long‐range couplings from N‐methyl groups to both α‐protons attached to that amide bond. (ii) An 1H, 13C‐COSY spectrum optimized for C,H‐long‐range couplings (J = 5 to 10 Hz) to the eleven CO groups again yields coupling to both α‐protons attached to that amide bond. Additionally these two experiments yield the assignment of N‐methyl protons and carbonyl C‐atoms. Normal and relayed 1H,13C‐COSY in both solvents have been applied to assign all C‐atoms via their directly attached and remote protons. An 1H,13C‐COLOC spectrum at 500 MHz in CDCl3, which uses H,C‐long‐range couplings confirms the assignment of all proton spin systems as well as the C‐signals of each individual amino acid. Ambiguities in the assignment of the C(δ)'s of MeLeu have thus been removed. An 1H,15N‐COSY spectrum enables the assignment of the 4 NH N‐atoms.

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“…In order to rule out such errors, 'H,I3C-HMBC experiments at natural abundance were performed, aiming at the detection of scalar through-bond interactions. In theory, the sequence can then be determined by connecting sequential (i-1 ,i) I3C' peptide-bond carbonyl carbon atoms via heteronuclear scalar through-bond sequential "C:-,-' HY and ' T -, -' W and intra-residue 'H:-'T: and ' H:-"C: connectivities as detected by 'H,'T-HMBC spectroscopy (Wynants et al, 1984;Bax and Marion, 1988;Hansen, 1991 ;Zimmermann et al, 1993), and homonuclear scalar intraresidue ' H:-' H: connectivities as detected by 'H-COSY and TOCSY, according to Scheme 3. Typical absolute values of the involved coupling constants from Scheme 3 for peptides are: 1 Hz, ZJ('H:-''C3 = 4-7 Hz, and 3J('Hy-'H7) = 4-12 Hz (Bystrov, 1976).…”

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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Assignment of amino acids in peptides by correlation of α-hydrogen and carbonyl carbon-13 resonances

Cited by 38 publications

References 26 publications

Isolation of 5-methoxy-2,2-dimethyl-1-2H-benzopyran-6-propanoic acid methyl ester and characterization by two-dimensional nuclear magnetic resonance spectroscopy

Isolation of 5-methoxy-2,2-dimethyl-1-2H-benzopyran-6-propanoic acid methyl ester and characterization by two-dimensional nuclear magnetic resonance spectroscopy

Peptide conformations. Part 30. Assignment of the ¹H‐, ¹³C‐, and ¹⁵N‐NMR spectra of cyclosporin A in CDCl₃ and C₆D₆ by a combination of homo‐ and heteronuclear two‐dimensional techniques

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

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Assignment of amino acids in peptides by correlation of α-hydrogen and carbonyl carbon-13 resonances

Cited by 38 publications

References 26 publications

Isolation of 5-methoxy-2,2-dimethyl-1-2H-benzopyran-6-propanoic acid methyl ester and characterization by two-dimensional nuclear magnetic resonance spectroscopy

Isolation of 5-methoxy-2,2-dimethyl-1-2H-benzopyran-6-propanoic acid methyl ester and characterization by two-dimensional nuclear magnetic resonance spectroscopy

Peptide conformations. Part 30. Assignment of the 1H‐, 13C‐, and 15N‐NMR spectra of cyclosporin A in CDCl3 and C6D6 by a combination of homo‐ and heteronuclear two‐dimensional techniques

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

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Peptide conformations. Part 30. Assignment of the ¹H‐, ¹³C‐, and ¹⁵N‐NMR spectra of cyclosporin A in CDCl₃ and C₆D₆ by a combination of homo‐ and heteronuclear two‐dimensional techniques