The Thermodynamics and Kinetics of Collagen Folding

Bächinger, Hans Peter; Engel, Jürgen

doi:10.1002/9783527619498.ch30

Cited by 14 publications

(36 citation statements)

References 162 publications

(225 reference statements)

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“…The observed hysteresis suggests that the cis – trans isomerization of the amide bonds involved in the transition from the PPI to the PPII helix and vice versa is slow compared to the change in the temperature. Hysteresis effects are also known for other systems of biological relevance, for example the folding/unfolding of the collagen triple helix32–34 and α‐helical coiled‐coil structures of SNARE proteins35. We therefore sought to obtain a deeper understanding of the transition between the PPI and the PPII helix and fitted the experimental data with a newly developed kinetic model for the quantitative description of hysteresis effects (32; within this publication, the newly developed formalism for the quantitative description of hysteresis effects of even complex systems is applied to collagen) to determine thermodynamic and kinetic parameters that characterize this molecular switch.…”

Section: Resultsmentioning

confidence: 99%

Temperature‐induced transition between polyproline I and II helices: quantitative fitting of hysteresis effects

Kuemin

Engel

Wennemers

2010

Journal of Peptide Science

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The conformational properties of the polyproline I (PPI) helix of oligoprolines toward heating were examined. Oligoproline H-Pro(12)-NH(2) served as a model which adopts in n-PrOH a pronounced PPI conformation with all cis amide bonds, whereas a polyproline II (PPII) conformation with all trans amide bonds is predominant in pure aqueous buffer. CD spectroscopic studies revealed that a conformational change from the PPI to the PPII helix takes place upon heating and back to the PPI helix upon cooling. This conformational transition cycle is characterized by a strong hysteresis. With a quantitative fitting of the experimentally observed hysteresis loops by a newly developed iterative integration with different starting conditions, kinetic and thermodynamic parameters for the transition from the PPI to the PPII helical conformation were determined. The transition is as expected for cis-trans isomerizations of amide bonds comparatively slow (k = 0.003 s(-1) at 80 °C) and characterized by an activation energy E(a) of 81.1 ± 3.6 kJ mol(-1). Thermodynamically, the transition from the PPI to the PPII helix is characterized by a positive standard enthalpy (ΔH(0) = 33.5 ± 2.1 kJ min(-1)) and a positive standard entropy (ΔS(0) = 102 ± 6.6 J mol(-1) K(-1)).

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Section: Resultsmentioning

confidence: 99%

Temperature‐induced transition between polyproline I and II helices: quantitative fitting of hysteresis effects

Kuemin

Engel

Wennemers

2010

Journal of Peptide Science

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“…Another explanation is that there is probably a difference in hydration of the unfolded chains. The peptide with 3(S)Hyp could be more hydrated than the peptide with Pro in single chains, which would cause a decrease in the transition enthalpy and a decrease in the entropy of solvent water (4,1,4), (3,2,4), (3,3,3), (1,7,1) or (0, 9, 0), were analyzed by DSC at 2 mM peptide concentration in NaCl ⁄ P i . The excess heat capacity is shown as a function of temperature with a scanning rate of 0.5°CAEmin molecules.…”

Section: Discussionmentioning

confidence: 99%

“…It comprises three left-handed polyproline II-like helices with a Gly-Xaa-Yaa repeat. These form a right-handed super helix with a one-residue stagger [1,2]. The collagen triple helix has many unique properties.…”

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Effect of the ‐Gly‐3(S)‐hydroxyprolyl‐4(R)‐hydroxyprolyl‐ tripeptide unit on the stability of collagen model peptides

et al. 2008

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The collagen triple helix is probably the most abundant protein motif in the human body. It comprises three left-handed polyproline II-like helices with a Gly-Xaa-Yaa repeat. These form a right-handed super helix with a one-residue stagger [1,2]. The collagen triple helix has many unique properties. One of them is the requirement for many post-translational modifications to produce the final tissue form of the molecule [3]. In vertebrate collagens, most of the Pro residues in the Yaa position of the -Gly-Xaa-Yaarepeat sequence are nearly completely 4-hydroxylated to 4(R)-hydroxyproline [4(R)Hyp] by the enzyme prolyl 4-hydroxylase (EC 1.14.11.2). This modification in the Yaa position is strongly related to the stability of the collagen triple helix. Prolyl 4-hydroxylation also occurs in the Xaa position in invertebrates. In addition to prolyl 4(R)-hydroxylation, a small numbers of proline residues are modified to 3(S)-hydroxyproline [3(S)Hyp] [4,5] in many types of vertebrate collagens, such as types I, II, III, IV, V and X. Invertebrate collagens also contain 3(S)Hyp, for example interstitial and cuticle collagens of annelids [6], crab sub-cuticular In order to evaluate the role of 3(S)-hydroxyproline [3(S)-Hyp] in the triple-helical structure, we produced a series of model peptides with nine tripeptide units including 0-9 3(S)-hydroxyproline residues. The sequences are H-(Gly-Pro-4(R)Hyp) l -(Gly-3(S)Hyp-4(R)Hyp) m -(Gly-Pro-4(R)Hyp) n -OH, where (l, m, n) = (9, 0, 0), (4, 1, 4), (3, 2, 4), (3,3,3), (1, 7, 1) and (0, 9, 0). All peptides showed triple-helical CD spectra at room temperature and thermal transition curves. Sedimentation equilibrium analysis showed that peptide H-(Gly-3(S)Hyp-4(R)Hyp) 9 -OH is a trimer. Differential scanning calorimetry showed that replacement of Pro residues with 3(S)Hyp residues decreased the transition enthalpy, and the transition temperature increases by 4.5°C from 52.0°C for the peptide with no 3(S)Hyp residues to 56.5°C for the peptide with nine 3(S)Hyp residues. The refolding kinetics of peptides H-(Gly-3(S)Hyp-4(R)Hyp) 9 -OH, H-(Gly-Pro-4(R)Hyp) 9 -OH and H-(Gly-4(R)Hyp-4(R)Hyp) 9 -OH were compared, and the apparent reaction orders of refolding at 10°C were n = 1.5, 1.3 and 1.2, respectively. Replacement of Pro with 3(S)Hyp or 4(R)Hyp has little effect on the refolding kinetics. This result suggests that the refolding kinetics of collagen model peptides are influenced mainly by the residue in the Yaa position of the -Gly-Xaa-Yaa-repeated sequence. The experiments indicate that replacement of a Pro residue by a 3(S)Hyp residue in the Xaa position of the -Gly-Xaa-4(R)Hyp-repeat of collagen model peptides increases the stability, mainly due to entropic factors.Abbreviations CRTAP, cartilage-associated protein; DSC, differential scanning calorimetry; Hyp, hydroxyproline; P3H1, prolyl 3-hydroxylase 1.

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“…These data suggest that the rate‐limiting step switched from nucleation to propagation at high peptide concentrations (35). The propagation of the CTH in collagen peptides is rate limiting because of the need for isomerization of the cis‐prolyl residues to the trans state, a requirement for CTH formation (18,37,45).…”

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Folding Studies of pH‐Dependent Collagen Peptides

Lee¹,

Chmielewski²

2010

Chem Biol Drug Des

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Synthetic collagen model peptides containing carboxylate-modified hydroxyproline residues (P E ) have been shown to form a collagen triple helix at low pH (1). Based on the fact that P E -containing peptides unfold on demand by altering the pH, we investigated the folding kinetics of a series of these collagen-based peptides. Refolding data indicated that the introduction of P E residues within a peptide affects the stability and refolding of the collagen peptides through electrostatic interactions and steric hindrance. Moreover, the specific placement of the P E residues within the collagen peptides allowed for selective testing for possible nucleation sites at the termini or the center of the peptides. Specifically, the data demonstrated that collagen peptides may nucleate from either terminally or internally located repeating sequences of Pro-Hyp-Gly. These studies indicate that the P E residue may play a useful role as a tool for studying the folding mechanism of synthetic collagen peptides.Key words: collagen, peptide, pH-dependent, folding, nucleation Abbreviations: Hyp, hydroxyproline; CTH, collagen triple helix; NC, non-collagenous; CD, circular dichroism; TFA, trifluoroacetic acid. Received 29 October 2009, revised 18 November 2009 accepted for publication 18 November 2009Collagen is one of the most abundant proteins found in mammals and is a major component of tissue, bone and cartilage. The major structural motif found in fibril forming collagen is the collagen triple helix (CTH) (2). This CTH motif is composed of three chains, each forming a left-handed type II polyproline helix, that associate and are stabilized by interchain hydrogen bonding within the CTH (3-6). To accommodate close packing, the CTH requires a glycine (Gly) residue at every third position within the helix, thereby generating repeating sequences of Gly-Xaa-Yaa in various types of collagen. A high imino acid content is found in the Xaa and Yaa positions, such as proline (Pro) and hydroxyproline (Hyp), and these residues have also been shown to be necessary in stabilizing the CTH (7-10).In nature, collagen is synthesized at the membrane of the rough endoplasmic reticulum in a procollagen form containing N-and C-terminal non-collagenous globular propeptides [non-collagenous (NC) domains] flanking the triple helical (Gly-Xaa-Yaa) n central collagenous domain (2,11-13). The assembly of the NC C-terminal domains is necessary for proper chain selection, registration and nucleation to generate an aligned triple helix (14-16). Nucleation of triple helix at (Gly-Pro-Hyp) rich sequences within the (Gly-Xaa-Yaa) regions is initiated through the NC domain assembly and is propagated in a zipper-like manner (17) with cis-trans isomerization as the rate-limiting step (18). In collagen III and IV, folding of triple helix proceeds from the C-to N-terminus, while other types of collagen display a variance in the directionality of their folding and nucleation (18-26).Although controversy exists over the reversibility of collagen triple helix formation...

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The Thermodynamics and Kinetics of Collagen Folding

Cited by 14 publications

References 162 publications

Temperature‐induced transition between polyproline I and II helices: quantitative fitting of hysteresis effects

Temperature‐induced transition between polyproline I and II helices: quantitative fitting of hysteresis effects

Effect of the ‐Gly‐3(S)‐hydroxyprolyl‐4(R)‐hydroxyprolyl‐ tripeptide unit on the stability of collagen model peptides

Folding Studies of pH‐Dependent Collagen Peptides

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