In pre-sterol carrier protein 2 (SCP2) in solution the leader peptide 1-20 is flexibly disordered, and residues 21-143 adopt the same globular fold as in mature SCP2

F. E. Weber, Jim Dyer, F. López García, M. Werder, T. Szyperski, K. Wüthrich, H. Hauser

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Abstract

The preform of the rabbit sterol carrier protein 2 (pre-rSCP2) was cloned, the uniformly 15N-labelled protein expressed in Escherichia coli and studied by three-dimensional 15N-resolved nuclear magnetic resonance spectroscopy. In spite of its low solubility in aqueous solution of only ~ 0.3 mM, sequential 15N and 1H backbone resonance assignments were obtained for 105 out of the 143 residues. From comparison of the sequential and medium-range nuclear Overhauser effects (NOEs) in the two proteins, all regular secondary structures previously determined in mature human SCP2 (hSCP2) were also identified in pre-rSCP2. Near-identity of the backbone 15N and 1H chemical shifts and 1:1 correspondence of 24 long-range NOEs to backbone amide groups in the two proteins show that the residues 21-143 adopt the same globular fold in pre-rSCP2 and mature hSCP2. The N-terminal 20-residue leader peptide of pre-rSCP2 is flexibly disordered in solution and does not observably affect the conformation of the polypeptide segment 21-143.

Original languageEnglish
Pages (from-to)751-759
Number of pages9
JournalCellular and Molecular Life Sciences
Volume54
Issue number7
DOIs
StatePublished - 28 Aug 1998

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Protein Sorting Signals
Rabbits
Proteins
Amides
Solubility
Magnetic Resonance Spectroscopy
sterol carrier proteins
Escherichia coli
Peptides

Keywords

  • N-terminal leader peptide
  • Nuclear magnetic resonance
  • Protein expression
  • Protein structure
  • Sterol carrier protein 2

Cite this

@article{5afd56a41ef44aeeb81b1a3af431f005,
title = "In pre-sterol carrier protein 2 (SCP2) in solution the leader peptide 1-20 is flexibly disordered, and residues 21-143 adopt the same globular fold as in mature SCP2",
abstract = "The preform of the rabbit sterol carrier protein 2 (pre-rSCP2) was cloned, the uniformly 15N-labelled protein expressed in Escherichia coli and studied by three-dimensional 15N-resolved nuclear magnetic resonance spectroscopy. In spite of its low solubility in aqueous solution of only ~ 0.3 mM, sequential 15N and 1H backbone resonance assignments were obtained for 105 out of the 143 residues. From comparison of the sequential and medium-range nuclear Overhauser effects (NOEs) in the two proteins, all regular secondary structures previously determined in mature human SCP2 (hSCP2) were also identified in pre-rSCP2. Near-identity of the backbone 15N and 1H chemical shifts and 1:1 correspondence of 24 long-range NOEs to backbone amide groups in the two proteins show that the residues 21-143 adopt the same globular fold in pre-rSCP2 and mature hSCP2. The N-terminal 20-residue leader peptide of pre-rSCP2 is flexibly disordered in solution and does not observably affect the conformation of the polypeptide segment 21-143.",
keywords = "N-terminal leader peptide, Nuclear magnetic resonance, Protein expression, Protein structure, Sterol carrier protein 2",
author = "Weber, {F. E.} and Jim Dyer and {L{\'o}pez Garc{\'i}a}, F. and M. Werder and T. Szyperski and K. W{\"u}thrich and H. Hauser",
year = "1998",
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language = "English",
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pages = "751--759",
journal = "Cellular and Molecular Life Sciences",
issn = "1420-682X",
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}

In pre-sterol carrier protein 2 (SCP2) in solution the leader peptide 1-20 is flexibly disordered, and residues 21-143 adopt the same globular fold as in mature SCP2. / Weber, F. E.; Dyer, Jim; López García, F.; Werder, M.; Szyperski, T.; Wüthrich, K.; Hauser, H.

In: Cellular and Molecular Life Sciences, Vol. 54, No. 7, 28.08.1998, p. 751-759.

Research output: Contribution to journalArticleResearchpeer-review

TY - JOUR

T1 - In pre-sterol carrier protein 2 (SCP2) in solution the leader peptide 1-20 is flexibly disordered, and residues 21-143 adopt the same globular fold as in mature SCP2

AU - Weber, F. E.

AU - Dyer, Jim

AU - López García, F.

AU - Werder, M.

AU - Szyperski, T.

AU - Wüthrich, K.

AU - Hauser, H.

PY - 1998/8/28

Y1 - 1998/8/28

N2 - The preform of the rabbit sterol carrier protein 2 (pre-rSCP2) was cloned, the uniformly 15N-labelled protein expressed in Escherichia coli and studied by three-dimensional 15N-resolved nuclear magnetic resonance spectroscopy. In spite of its low solubility in aqueous solution of only ~ 0.3 mM, sequential 15N and 1H backbone resonance assignments were obtained for 105 out of the 143 residues. From comparison of the sequential and medium-range nuclear Overhauser effects (NOEs) in the two proteins, all regular secondary structures previously determined in mature human SCP2 (hSCP2) were also identified in pre-rSCP2. Near-identity of the backbone 15N and 1H chemical shifts and 1:1 correspondence of 24 long-range NOEs to backbone amide groups in the two proteins show that the residues 21-143 adopt the same globular fold in pre-rSCP2 and mature hSCP2. The N-terminal 20-residue leader peptide of pre-rSCP2 is flexibly disordered in solution and does not observably affect the conformation of the polypeptide segment 21-143.

AB - The preform of the rabbit sterol carrier protein 2 (pre-rSCP2) was cloned, the uniformly 15N-labelled protein expressed in Escherichia coli and studied by three-dimensional 15N-resolved nuclear magnetic resonance spectroscopy. In spite of its low solubility in aqueous solution of only ~ 0.3 mM, sequential 15N and 1H backbone resonance assignments were obtained for 105 out of the 143 residues. From comparison of the sequential and medium-range nuclear Overhauser effects (NOEs) in the two proteins, all regular secondary structures previously determined in mature human SCP2 (hSCP2) were also identified in pre-rSCP2. Near-identity of the backbone 15N and 1H chemical shifts and 1:1 correspondence of 24 long-range NOEs to backbone amide groups in the two proteins show that the residues 21-143 adopt the same globular fold in pre-rSCP2 and mature hSCP2. The N-terminal 20-residue leader peptide of pre-rSCP2 is flexibly disordered in solution and does not observably affect the conformation of the polypeptide segment 21-143.

KW - N-terminal leader peptide

KW - Nuclear magnetic resonance

KW - Protein expression

KW - Protein structure

KW - Sterol carrier protein 2

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