HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

J. Biol. Chem., Vol. 279, Issue 36, 38072-38078, September 3, 2004

This Article Full Text Full Text (PDF) All Versions of this Article: 279/36/38072    most recent M402953200v1 Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Mizuno, K. Articles by Bächinger, H. P. Search for Related Content PubMed PubMed Citation Articles by Mizuno, K. Articles by Bächinger, H. P. Social Bookmarking                      What's this?

Hydroxylation-induced Stabilization of the Collagen Triple Helix

ACETYL-(GLYCYL-4(R)-HYDROXYPROLYL-4(R)-HYDROXYPROLYL)₁₀-NH₂ FORMS A HIGHLY STABLE TRIPLE HELIX^*

Kazunori Mizuno, Toshihiko Hayashi, David H. Peyton¶, and Hans Peter Bächinger||

From the Department of Biochemistry and Molecular Biology, Oregon Health & Science University, and Shriners Hospital for Children, Research Department, Portland, Oregon 97239, the ^¶Department of Chemistry, Portland State University, Portland, Oregon 97201, and the Faculty of Pharmaceutical Sciences, Teikyo Heisei University, Ichihara, 290-0193, Japan

The collagen triple helix is one of the most abundant protein motifs in animals. The structural motif of collagen is the triple helix formed by the repeated sequence of -Gly-Xaa-Yaa-. Previous reports showed that H-(Pro-4(R)Hyp-Gly)₁₀-OH (where `4(R)Hyp' is (2S,4R)-4-hydroxyproline) forms a trimeric structure, whereas H-(4(R)Hyp-Pro-Gly)₁₀-OH does not form a triple helix. Compared with H-(Pro-Pro-Gly)₁₀-OH, the melting temperature of H-(Pro-4(R)Hyp-Gly)₁₀-OH is higher, suggesting that 4(R)Hyp in the Yaa position has a stabilizing effect. The inability of triple helix formation of H-(4(R)Hyp-Pro-Gly)₁₀-OH has been explained by a stereoelectronic effect, but the details are unknown. In this study, we synthesized a peptide that contains 4(R)Hyp in both the Xaa and the Yaa positions, that is, Ac-(Gly-4(R)Hyp-4(R)Hyp)₁₀-NH₂ and compared it to Ac-(Gly-Pro-4(R)Hyp)₁₀-NH₂, and Ac-(Gly-4(R)Hyp-Pro)₁₀-NH₂. Ac-(Gly-4(R)Hyp-4(R)Hyp)₁₀-NH₂ showed a polyproline II-like circular dichroic spectrum in water. The thermal transition temperatures measured by circular dichroism and differential scanning calorimetry were slightly higher than the values measured for Ac-(Gly-Pro-4(R)Hyp)₁₀-NH₂ under the same conditions. For Ac-(Gly-4(R)Hyp-4(R)Hyp)₁₀-NH₂, the calorimetric and the van't Hoff transition enthalpy H were significantly smaller than that of Ac-(Gly-Pro-4(R)Hyp)₁₀-NH₂. We postulate that the denatured states of the two peptides are significantly different, with Ac-(Gly-4(R)Hyp-4(R)Hyp)₁₀-NH₂ forming a more polyproline II-like structure instead of a random coil. Two-dimensional nuclear Overhauser effect spectroscopy suggests that the triple helical structure of Ac-(Gly-4(R)Hyp-4(R)Hyp)₁₀-NH₂ is more flexible than that of Ac-(Gly-Pro-4(R)Hyp)₁₀-NH₂. This is confirmed by the kinetics of amide ¹H exchange with solvent deuterium of Ac-(Gly-4(R)Hyp-4(R)Hyp)₁₀-NH₂, which is faster than that of Ac-(Gly-Pro-4(R)Hyp)₁₀-NH₂. The higher transition temperature of Ac-(Gly-4(R)Hyp-4(R)Hyp)₁₀-NH₂, can be explained by the higher trans/cis ratio of the Gly-4(R)Hyp peptide bonds than that of the Gly-Pro bonds, and this ratio compensates for the weaker interchain hydrogen bonds.

Received for publication, March 17, 2004 , and in revised form, June 14, 2004.

^* This work was supported by a grant from the Shriners Hospital for Children. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

^|| To whom correspondence should be addressed: Research Dept., Shriners Hospital for Children, 3101 SW Sam Jackson Park Rd., Portland, OR 97239. Tel.: 503-221-3433; Fax: 503-221-3451; E-mail: hpb{at}shcc.org.

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				R. Improta, R. Berisio, and L. Vitagliano Contribution of dipole-dipole interactions to the stability of the collagen triple helix Protein Sci., May 1, 2008; 17(5): 955 - 961. [Abstract] [Full Text] [PDF]

				M. Schumacher, K. Mizuno, and H. P. Bachinger The Crystal Structure of the Collagen-like Polypeptide (Glycyl-4(R)-hydroxyprolyl-4(R)-hydroxyprolyl)9 at 1.55 A Resolution Shows Up-puckering of the Proline Ring in the Xaa Position J. Biol. Chem., May 27, 2005; 280(21): 20397 - 20403. [Abstract] [Full Text] [PDF]