<sup>13</sup>C nuclear magnetic resonance study of the triple helix ⇌ random‐coil transition of (prolylprolyglycyl)<sub>10</sub>

Blasi, R. Di; Verdini, A. S.

doi:10.1002/bip.1979.360180318

Cited by 13 publications

(4 citation statements)

References 10 publications

(1 reference statement)

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“…The T1 values of both of the carbonyl carbons were then averaged. The 13C NMR peak assignments of the carbonyl carbons attributed to the amide and the carboxylic acid groups in the NMwA molecule and to the amino acid residues in the (PPG)s molecule were done as described in the literature (Di Blasi and Verdini, 1979;Nishiyama et al, 1995Nishiyama et al, , 1998Nishiyama et al, , 2000Suzuki et al, 1998).…”

Section: Preparation Of N-methacryloyl-o-amino Acidsmentioning

confidence: 99%

Adhesion of N-Methacryloyl-ω-Amino Acid Primers to Collagen Analyzed by 13C NMR

et al. 2001

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Previously, we reported that the strength of the interaction between N-methacryloyl-omega-amino acid (NMomegaA) primers and dentinal collagen exhibited a strong correlation with the bond strength of the resin to etched dentin. To determine the pertinent functional groups of the amino acid residues in the dentinal collagen, to which the amide and/or the carboxylic acid groups of the NMomegaAs are adsorbed, we used 13C NMR techniques--primarily through the observation of spin-lattice relaxation times, Ti--to investigate the adsorption characteristics resulting from the interaction of NMomegaAs with a model oligopeptide for collagen, (PPG)5. The addition of NMomegaAs to a collagenous solution resulted in a decrease in the T1 values of the carbonyl carbons attributed to the carboxylic acid of the C-terminal Gly and to the third amide of the N-terminal Pro residues in the (PPG)5 molecule, thus reflecting the formation of hydrogen-bonded interactions.

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Section: Preparation Of N-methacryloyl-o-amino Acidsmentioning

confidence: 99%

Adhesion of N-Methacryloyl-ω-Amino Acid Primers to Collagen Analyzed by 13C NMR

et al. 2001

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“…Using small collagen peptides and polypeptides such as the a 1-CB2 fragment of rat skin collagen, ID1 13C assignments of aliphatic resonances in the random coil state have been obtained, and the mobility of residues in the coil and helix forms has been examined (Torchia & Lyerla, 1974;Torchia et al, 1975). NMR has also proved useful in detecting and quantitating the cis form of X-Pro (Hyp) peptide bonds in collagen (Roques et al, 1977;Di Blasi & Verdini, 1979;Sarkar et al, 1984;Torchia et al, 1985) and in studying single-chain X-Y-Gly repeating peptides (Mayo et al, 1991).…”

mentioning

confidence: 99%

Two-dimensional NMR assignments and conformation of (Pro-Hyp-Gly)10 and a designed collagen triple-helical peptide

et al. 1993

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Homonuclear and heteronuclear 2D NMR methods are used to study two triple-helical peptides. One peptide, (POG)10, is considered to be the most stable prototype of a triple helix. The second peptide, (POG)3ITGARGLAGPOG(POG)3 (denoted T3-785), was designed to model an imino acid poor region of collagen and contains 12 residues from near the unique collagenase cleavage site in type III collagen. Both peptides associated as trimers, with melting temperatures of 60 degrees C for (POG)10 and 25 degrees C for the T3-785 peptide. Sequence-specific assignments were made for a tripeptide unit POG in (POG)10, and 80% of the POG triplets are found to be in an equivalent environment. In T3-785, with nonrepeating X-Y-Gly units incorporated in the sequence, the three chains of the homotrimer can be distinguished from one another by NMR. The solution conformation of (POG)10 is very similar to the model derived from X-ray fiber diffraction data, although the peptide contains less ordered regions at the peptide ends. In the trimer from of T3-785, the central residues of the three chains are closely packed, and the data are consistent with a triple-helical model with a one-residue stagger of three parallel chains. For T3-785, in contrast to (POG)10, there are also resonances from a less ordered form, which are probably due to the presence of a small amount of monomer. The similarity of the backbone conformations of T3-785 and (POG)10 suggests that an alternative conformation is not present in the imino acid poor region.

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“…In view of these observations and the known propensity of -Pro-Gly-segments to take up a folded conformation, we might expect the nonhelical form of (Pro-Pro-Gly)n polypeptides to contain several folded segments that resemble the (3-turn (type I, II, or III, depending on the particular sets of 4, 4i angles for the proline and glycine residues); however, such segments cannot be additionally stabilized by the intramolecular hydrogen bonding (due to lack of hydrogen atom on the imide nitrogen). [On the basis of available NMR data (24), the cis isomer is unlikely to be significantly populated as a result of heating aqueous solutions of (Pro-Pro-Gly)1o.] The observed CD spectrum of (ProPro-Gly)10 in the incubation mixture prior to the onset of the hydroxylation process may be taken to correspond to the nonhelical conformation of the polypeptide and may serve as an approximation to that of the unhydroxylated procollagen.…”

Section: Discussionmentioning

confidence: 99%

Conformational implications of enzymatic proline hydroxylation in collagen.

Chopra¹,

Ananthanarayanan²

1982

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

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In 1979 it was proposed that prolyl hydroxylase (prolyl-glycyl-peptide,2-oxoglutarate:oxygen oxidoreductase, EC 1. 14.11.2) recognizes the 3-turn conformation in nascent procollagen chains and that the hydroxylation process involves a conformational change resulting in "straightening" of the (turn segments into the linear triple-helical conformation of native collagen. We present experimental data that verify both these postulates. The following peptides were synthesized and studied for their conformation and interaction with prolyl hydroxylase: tBoc-Pro-GlyAla-OH, tBoc-Pro-Gly-Val-OH, tBoc-Gly-Val-Pro-Gly-Val-OH, and tBoc-Pro-DAla-Ala-OH. Spectral data showed that these peptides preferred a (-turn conformation. All of them acted as inhibitors of the enzyme; the pentapeptide also acted as a substrate. To mimic the biosynthetic event, a collagen model polypeptide, (Pro-Pro-Gly)10, was incubated at 37C with purified prolyl hydroxylase and the necessary cosubstrates and cofactors at pH 7.8. A progressive change from the initially nonhelical to the triplehelical conformation, as monitored by CD spectra and gel filtration, occurred during the course of proline hydroxylation. In addition to leading to increased thermal stability of the triple-helical conformation in (Pro-Pro-Gly)10 and (Pro-Pro-Gly)5, the enzymatic incorporation of the hydroxyproline residues was found to enable these polypeptides to fold into this conformation faster than the unhydroxylated counterparts. These conformational implications ofproline hydroxylation in collagen may also be of use in the study of the complement subcomponent Clq and of acetylcholine esterase which contain collagen-like regions in them.The post-translational hydroxylation of selected proline residues by prolyl hydroxylase (prolyl-glycyl-peptide,2-oxoglutarate:oxygen oxidoreductase, EC 1.14.11.2) is a crucial event in the biosynthesis of collagen. The resulting hydroxyproline residues are essential for the stability of the triple-helical conformation of collagen at body temperature (1). In regard to the conformational aspects of the enzymatic hydroxylation process, two questions needed to be answered: (a) What is the preferred conformation of the peptide substrate? and (b) What is the conformational consequence of proline hydroxylation? Based on experimental data and theoretical considerations, it was proposed (2) that prolyl hydroxylase recognizes the (3-turn conformation (3) formed at the -Pro-Gly-segments in the nascent procollagen chains and that the hydroxylation process results in "straightening" of these segments into the rigid conformation necessary for the subsequent association of these chains in the triple-helical conformation of native collagen.In order to test these postulates in a direct manner, we undertook (a) the synthesis of simple peptides which were expected to prefer the (3-turn conformation to study their interactions with prolyl hydroxylase and (b) MATERIALS AND METHODS Enzyme. Prolyl hydroxylase from 13-day chicken embryos was purified to hom...

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¹³C nuclear magnetic resonance study of the triple helix ⇌ random‐coil transition of (prolylprolyglycyl)₁₀

Cited by 13 publications

References 10 publications

Adhesion of N-Methacryloyl-ω-Amino Acid Primers to Collagen Analyzed by 13C NMR

Adhesion of N-Methacryloyl-ω-Amino Acid Primers to Collagen Analyzed by 13C NMR

Two-dimensional NMR assignments and conformation of (Pro-Hyp-Gly)10 and a designed collagen triple-helical peptide

Conformational implications of enzymatic proline hydroxylation in collagen.

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13C nuclear magnetic resonance study of the triple helix ⇌ random‐coil transition of (prolylprolyglycyl)10

Cited by 13 publications

References 10 publications

Adhesion of N-Methacryloyl-ω-Amino Acid Primers to Collagen Analyzed by 13C NMR

Adhesion of N-Methacryloyl-ω-Amino Acid Primers to Collagen Analyzed by 13C NMR

Two-dimensional NMR assignments and conformation of (Pro-Hyp-Gly)10 and a designed collagen triple-helical peptide

Conformational implications of enzymatic proline hydroxylation in collagen.

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¹³C nuclear magnetic resonance study of the triple helix ⇌ random‐coil transition of (prolylprolyglycyl)₁₀