Crystal structure of a collagen-like polypeptide with repeating sequence Pro-Hyp-Gly at 1.4 � resolution: Implications for collagen hydration

Berisio, Rita; Vitagliano, Luigi; Mazzarella, L.; Zagari, Adriana

doi:10.1002/1097-0282(2000)56:1<8::aid-bip1037>3.0.co;2-w

Cited by 102 publications

(96 citation statements)

References 26 publications

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“…Standard peptides containing only Gly-Pro-Hyp repeating sequences, (POG) 10 and (GPO) 8 , are considered as controls. 20,21,23 Peptides containing interruptions of seven residue or fewer in length are all fully soluble in phosphate-buffered saline (PBS), pH 7, at a concentration of 1 mg mL À1 at low temperature. The two peptides with the longest interruptions (m ¼ 8 and 9) showed some aggregation under these conditions, and a procedure of heating, cooling, and centrifugation was used to prepare a solution for physical chemical characterization (final concentration of 1 mg mL À1 ).…”

Section: Resultsmentioning

confidence: 99%

Interruptions in the collagen repeating tripeptide pattern can promote supramolecular association

Hwang¹,

Thiagarajan²,

Parmar³

et al. 2010

Protein Science

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The standard collagen triple-helix requires a perfect (Gly-Xaa-Yaa) n sequence, yet all nonfibrillar collagens contain interruptions in this tripeptide repeating pattern. Defining the structural consequences of disruptions in the sequence pattern may shed light on the biological role of sequence interruptions, which have been suggested to play a role in molecular flexibility, collagen degradation, and ligand binding. Previous studies on model peptides with 1-and 4-residue interruptions showed a localized perturbation within the triple-helix, and this work is extended to introduce natural collagen interruptions up to nine residue in length within a fixed (Gly-Pro-Hyp) n peptide context. All peptides in this set show decreases in triple-helix content and stability, with greater conformational perturbations for the interruptions longer than five residue. The most stable and least perturbed structure is seen for the 5-residue interruption peptide, whose sequence corresponds to a Gly to Ala missense mutation, such as those leading to collagen genetic diseases. The triple-helix peptides containing 8-and 9-residue interruptions exhibit a strong propensity for self-association to fibrous structures. In addition, a small peptide modeling only the 9-residue sequence within the interruption aggregates to form amyloid-like fibrils with antiparallel b-sheet structure. The 8-and 9-residue interruption sequences studied here are predicted to have significant cross-b aggregation potential, and a similar propensity is reported for 10% of other naturally occurring interruptions. The presence of amyloidogenic sequences within or between triple-helix domains may play a role in molecular association to normal tissue structures and could participate in observed interactions between collagen and amyloid.

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Section: Resultsmentioning

confidence: 99%

Interruptions in the collagen repeating tripeptide pattern can promote supramolecular association

Hwang¹,

Thiagarajan²,

Parmar³

et al. 2010

Protein Science

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“…22 For example, the backbone dihedral angles for 2 are intermediate between those of 1 and AcFlpOMe (7), and are favorable for triple helix formation. 23,24 13 C NMR chemical shifts can report on electron withdrawal by pendant function groups. 25 The relative electron-withdrawing ability of the hydroxyl, acetoxyl, and trifluoroacetoxyl groups in 1-6 were assessed by comparing the 13 C NMR chemical shifts of their C γ atoms with those of AcProOMe, AcFlpOMe (7), and AcflpOMe (8).…”

Section: Resultsmentioning

confidence: 99%

O‐acylation of hydroxyproline residues: Effect on peptide‐bond isomerization and collagen stability

et al. 2004

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In collagen, strands of the sequence XaaYaaGly form a triple‐helical structure. The Yaa residue is often (2S,4R)‐4‐hydroxyproline (Hyp). The inductive effect of the hydroxyl group of Hyp residues greatly increases collagen stability. Here, electron withdrawal by the hydroxyl group in Hyp and its 4S diastereomer (hyp) is increased by the addition of an acetyl group or trifluoroacetyl group. The crystalline structures of AcHyp[C(O)CH3]OMe and Achyp[C(O)CH3]OMe are similar to those of AcHypOMe and AcProOMe, respectively. The O‐acylation of AcHypOMe and AchypOMe increases the 13C chemical shift of its Cγ atom: AcHyp[C(O)CF3]OMe ≅ Achyp[C(O)CF3]OMe > AcHyp[C(O)CH3]OMe ≅ Achyp[C(O)CH3]OMe ≥ AcHypOMe ≅ AchypOMe. This increased inductive effect is not apparent in the thermodynamics or kinetics of amide bond isomerization. Despite apparently unfavorable steric interactions, (ProHypGly)10, which is O‐acylated with 10 acetyl groups, forms a triple helix that has intermediate stability: (ProHypGly)10 > {ProHyp[C(O)CH3]Gly}10 ≫ (ProProGly)10. Thus, the benefit to collagen stability endowed by the hydroxyl group of Hyp residues is largely retained by an acetoxyl group. © 2004 Wiley Periodicals, Inc. Biopolymers (Pept Sci), 2005

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“…The data contained both very strong and very weak reflections, as had been observed previously for crystalline triple helices (58). Data were processed with HKL2000 (59), and initial phases were obtained by molecular replacement by using a truncated model of a crystalline ½ðProProGlyÞ 10 3 triple-helix structure determined by Berisio, Vitagliano, Mazzarella, and Zagari (Protein Data Bank entry 1k6f) (35).…”

Section: Methodsmentioning

confidence: 99%

Stereoelectronic and steric effects in side chains preorganize a protein main chain

Shoulders¹,

Satyshur²,

Forest³

et al. 2009

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

153

142

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Preorganization is shown to endow a protein with extraordinary conformational stability. This preorganization is achieved by installing side-chain substituents that impose stereoelectronic and steric effects that restrict main-chain torsion angles. Replacing proline residues in ðProProGlyÞ 7 collagen strands with 4-fluoroproline and 4-methylproline leads to the most stable known triple helices, having T m values that are increased by >50°C. Differential scanning calorimetry data indicate an entropic basis to the hyperstability, as expected from an origin in preorganization. Structural data at a resolution of 1.21 Å reveal a prototypical triple helix with insignificant deviations to its main chain, even though 2∕3 of the residues are nonnatural. Thus, preorganization of a main chain by subtle changes to side chains can confer extraordinary conformational stability upon a protein without perturbing its structure.collagen triple helix | nonnatural amino acid | preorganization | protein stability | x-ray crystallography T he three-dimensional structures of proteins are stable because of a delicate balance of forces (1). Increases in the conformational stability of a protein-desirable in many contexts (2)-can be realized by enhancing the hydrophobic effect (3, 4), introducing or shielding a hydrogen bond (5) or electrostatic interaction (6-8), or introducing a disulfide crosslink (9-11) or metal ion-binding site (12). These strategies are often frustrated by enthalpy-entropy compensation, whereby enthalpic stabilization is offset entirely by entropic destabilization, or vice versa (13). Moreover, the strategies often lack subtlety and can lead to a misshapen and hence dysfunctional protein.As elaborated by Cram (14), the principle of preorganization states that, "the more highly hosts and guests are organized for binding and low solvation prior to their complexation, the more stable will be their complexes." In a typical manifestation, a conformational constraint is imposed upon a receptor or ligand during its chemical synthesis. This principle can also yield biopolymers with increased conformational stability (Fig. 1). For example, the stability of a DNA duplex correlates with the helicity of its single strands (15, 16), and a bicyclic ("locked") derivative of ribose enhances that stability (17, 18). The stability of β-turns within protein structures can be increased by incorporating Dproline (19) and certain β-amino acids (20) that bias the conformations occupied by the unfolded polypeptide. Likewise, proteins containing a cis-peptide bond can be stabilized by constrained amides or isosteres (21-23). The utility of these substitutions is limited, however, by the difficulty of modifying the backbone of proteins as well as concomitant effects on structure and function (24-27).We suspected that alterations to proline residues could provide a particularly incisive means to preorganize the conformation of a polypeptide chain. In classic work, Matthews and coworkers substituted proline, which is the least flexible residu...

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Crystal structure of a collagen-like polypeptide with repeating sequence Pro-Hyp-Gly at 1.4 � resolution: Implications for collagen hydration

Cited by 102 publications

References 26 publications

Interruptions in the collagen repeating tripeptide pattern can promote supramolecular association

Interruptions in the collagen repeating tripeptide pattern can promote supramolecular association

O‐acylation of hydroxyproline residues: Effect on peptide‐bond isomerization and collagen stability

Stereoelectronic and steric effects in side chains preorganize a protein main chain

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