A Peptide Corresponding to Residues Asp177 to Asn208 of Human Cyclin A Forms an α-Helix

doi:10.1006/bbrc.1998.9828

Biochemical and Biophysical Research Communications

Volume 253, Issue 3, 30 December 1998, Pages 621-627

https://doi.org/10.1006/bbrc.1998.9828 Get rights and content

Abstract

Cyclins are essential activators of eukaryotic cell cycle-regulating enzymes called cyclin-dependent kinases (CDKs). The binding of cyclins to CDKs is mediated by a structural motif comprising a five-helix bundle called the cyclin fold and an additional helix (the N-terminal α-helix) located N-terminal to the cyclin fold. In this work, we examine, using CD and NMR spectroscopy, the structure of a 32-residue synthetic peptide derived from the segment (Asp177 to Asn208) corresponding to the N-terminal α-helix of human cyclin A. CD spectroscopic analysis of the peptide revealed that trifluoroethanol (TFE) can induce the peptide to assume a stable α-helix conformation. Two-dimensional¹H NMR spectroscopy showed that the α-helix is formed by the Asp181 to Cys193 segment of the peptide. The α-helical structure of the peptide in the TFE/H₂O cosolvent was found to be identical to that in the crystal structure of intact cyclin A. Taken together, these results suggest that the N-terminal α-helix of cyclins may exist as an independent structural unit that plays essential functional roles in activating CDKs.

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^☆: Abbreviations used: CD, circular dichroism; CDK, cyclin-dependent kinase; DTT, dithiothreitol; HMQC, heteronuclear multiple quantum correlation; NOE, nuclear Overhauser enhancement; NOESY, NOE spectroscopy; TFE, 2,2,2-trifluoroethanol; TOCSY, total correlation spectroscopy; [Θ]₂₂₂, mean ellipticity at 222 nm

¹: To whom correspondence should be addressed. E-mail:[email protected].

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Regular ArticleA Peptide Corresponding to Residues Asp177 to Asn208 of Human Cyclin A Forms an α-Helix☆

Abstract

Structure

J. Magn. Reson.

J. Magn. Reson.

J. Magn. Reson.

Methods Enzymol.

J. Mol. Biol.

Nature

Nature

Nature

Nat. Struct. Biol.

EMBO J.

Nat. Struct. Biol.

Cell

Regular Article
A Peptide Corresponding to Residues Asp177 to Asn208 of Human Cyclin A Forms an α-Helix☆