Investigation of the formation of triple helices between (pro‐pro‐gly)<sub><i>n</i></sub>'s of different degrees of polymerization using gel filtration

Kobayashi, Yuji; Suezaki, Yukio; Inouye, Katsuhiko; Tanaka, K.; Kakiuchi, Kinji; Kyōgoku, Yoshimasa

doi:10.1002/bip.1977.360160203

Cited by 3 publications

(7 citation statements)

References 17 publications

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“…From the characteristics of the CD spectra, from the gel filtration patterns, and from the earlier studies by other workers (13,14), we conclude that the materials eluting as trimers in the gel filtration are in the triple-helical conformation similar but not necessarily identical to that of native collagen. The possi- bility of mismatch among the constituent polypeptide chains appears to be small, if not absent (13,14).…”

Section: Conformation Of Synthetic Peptides and Theirsupporting

confidence: 60%

“…The limited number ofearlier studies (16,17) on the interaction of oligopeptides did not address themselves to the conformation of the peptides used. In the present study, the tripeptides tBoc-Pro-Gly-Ala-OH and tBoe-Pro-Gly-Val-OH were found to exhibit the type II ,&tum conformation ( §), and tBoc-Pro-DAla-Ala-OH was found (8) to take up the Venkatachalam type 13 (a subclass ofthe type II),&turn. The pentapeptide tBoc-Gly-Val-Pro-Gly-Val-OH was chosen on the basis of the observed (-turn preferences of similar sequences in tropoelastin model peptides (18).…”

Section: Discussionmentioning

confidence: 46%

“…In water, the intramolecular hydrogen bonding weakened, leading to a disordered structure. tBoc-Pro-D-Ala-Ala was found (8) to take up the Venkatachalam type 13 8-turn which was predicted for peptides with the L-D sequence (11). This conformation was found in both nonaqueous solvents and water.…”

Section: Conformation Of Synthetic Peptides and Theirmentioning

confidence: 73%

“…The possi- bility of mismatch among the constituent polypeptide chains appears to be small, if not absent (13,14). The data on the relative stabilities of (Pro-Pro-Gly)5 and (Pro-Pro-Gly)j0 and their hydroxylated counterparts are presented in Fig.…”

Section: Conformation Of Synthetic Peptides and Theirmentioning

confidence: 96%

“…The state ofassociation of(Pro-Pro-Gly)l0 after the enzymatic proline hydroxylation was determined by separating the components of the reaction mixture by gel filtration on Sephadex G-50 at 40C and comparing the elution profile of the hydroxylated polypeptide with that of the control (13,14). The data (not shown) revealed that the former had a relatively much larger proportion of the trimeric relative to monomeric species than the latter.…”

Section: Conformation Of Synthetic Peptides and Theirmentioning

confidence: 99%

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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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Section: Conformation Of Synthetic Peptides and Theirsupporting

confidence: 60%

Section: Discussionmentioning

confidence: 46%

Section: Conformation Of Synthetic Peptides and Theirmentioning

confidence: 73%

Section: Conformation Of Synthetic Peptides and Theirmentioning

confidence: 96%

Section: Conformation Of Synthetic Peptides and Theirmentioning

confidence: 99%

See 3 more Smart Citations

Conformational implications of enzymatic proline hydroxylation in collagen.

Chopra¹,

Ananthanarayanan²

1982

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

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Association of poly-(L-lysine) homologues in sodium carbonate solution

Kakiuchi

Tsuboi

1990

Colloid & Polymer Sci

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The molecular weights of isopoly(L-lysine), poly(L-ornithine), and poly(L-a, }~diaminobutylic acid), the homologues of poly (L-lysine), were determined by the sedimentation equilibrium method in aqueous solutions of 1.0 M NaC1 or 0.1 M Na2CO3. In every sample the molecular weights in the presence of carbonate ions was twice that in NaC1 solution. In a previous paper we reported that poly(L-lysine) behaved as a dimer at concentrations higher than 0.4 g/dl in the presence of carbonate ions and as a monomer in dilute solution, and these two forms were related by a monomer-dimer equilibrium. The homologues did not have a monomer-dimer equilibrium relationship under the conditions of the measurements that we carried out. The CD spectrum of isopoly(L-lysine) in water showed a uniform increase with a decrease in the wave length in the presence of carbonate ions. However, in the alkaline region in NaOH solution, the spectrum changed and a small minimum at 212 nm was found. When additional carbonate ions were added a large minimum at 205 nm was observed. This result can be explained by a change in the conformation from a random coil to a regular structure. We could not compare isopoly(L-lysine) with other polypeptides, because it does not have peptide bonds. The CD spectra of poly(L-ornithine) and poly(L-a, ~-diaminobutylic acid) in NaOH or Na2CO3 solutions showed only slightly regular structures. It was also confirmed that the dimer-structures of the poly(L-lysine) homologues do not have regular structures.

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Cooperative conformational transitions in linear macromolecules undergoing chiral perturbations

Teŕamoto

2001

Progress in Polymer Science

103

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Investigation of the formation of triple helices between (pro‐pro‐gly)_n's of different degrees of polymerization using gel filtration

Cited by 3 publications

References 17 publications

Conformational implications of enzymatic proline hydroxylation in collagen.

Conformational implications of enzymatic proline hydroxylation in collagen.

Association of poly-(L-lysine) homologues in sodium carbonate solution

Cooperative conformational transitions in linear macromolecules undergoing chiral perturbations

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Investigation of the formation of triple helices between (pro‐pro‐gly)n's of different degrees of polymerization using gel filtration

Cited by 3 publications

References 17 publications

Conformational implications of enzymatic proline hydroxylation in collagen.

Conformational implications of enzymatic proline hydroxylation in collagen.

Association of poly-(L-lysine) homologues in sodium carbonate solution

Cooperative conformational transitions in linear macromolecules undergoing chiral perturbations

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Product

Resources

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Investigation of the formation of triple helices between (pro‐pro‐gly)_n's of different degrees of polymerization using gel filtration