Evolutionary bridges to new protein folds: Design of C-terminal Cro protein chameleon sequences

William J. Anderson, Laura O. Van Dorn, Wendy M. Ingram, Matthew Hj Cordes

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

Regions of amino-acid sequence that are compatible with multiple folds may facilitate evolutionary transitions in protein structure. In a previous study, we described a heuristically designed chameleon sequence (SASF1, structurally ambivalent sequence fragment 1) that could adopt either of two naturally occurring conformations (α-helical or β-sheet) when incorporated as part of the C-terminal dimerization subdomain of two structurally divergent transcription factors, P22 Cro and λ Cro. Here we describe longer chameleon designs (SASF2 and SASF3) that in the case of SASF3 correspond to the full C-terminal half of the ordered region of a P22 Cro/λ Cro sequence alignment (residues 34-57). P22-SASF2 and λWDD-SASF2 show moderate thermal stability in denaturation curves monitored by circular dichroism (Tm values of 46 and 55°C, respectively), while P22-SASF3 and λWDD-SASF3 have somewhat reduced stability (Tm values of 33 and 49°C, respectively). 13C and 1H NMR secondary chemical shift analysis confirms two C-terminal α-helices for P22-SASF2 (residues 36-45 and 54-57) and two C-terminal β-strands for λWDD-SASF2 (residues 4045 and 5052), corresponding to secondary structure locations in the two parent sequences. Backbone relaxation data show that both chameleon sequences have a relatively well-ordered structure. Comparisons of 15N- 1H correlation spectra for SASF2 and SASF3-containing proteins strongly suggest that SASF3 retains the chameleonism of SASF2. Both Cro C-terminal conformations can be encoded in a single sequence, showing the plausibility of linking different Cro folds by smooth evolutionary transitions. The N-terminal subdomain, though largely conserved in structure, also exerts an important contextual influence on the structure of the C-terminal region.

Original languageEnglish (US)
Pages (from-to)765-771
Number of pages7
JournalProtein Engineering, Design and Selection
Volume24
Issue number9
DOIs
StatePublished - Sep 2011

Fingerprint

Lizards
Conformations
Proteins
Denaturation
Transcription factors
Dimerization
Chemical shift
Dichroism
Amino acids
Thermodynamic stability
Transcription Factors
Sequence Alignment
Nuclear magnetic resonance
Circular Dichroism
Amino Acids
Amino Acid Sequence
Hot Temperature

Keywords

  • chameleon sequence
  • NMR spectroscopy
  • protein design

ASJC Scopus subject areas

  • Biochemistry
  • Biotechnology
  • Bioengineering
  • Molecular Biology

Cite this

Evolutionary bridges to new protein folds : Design of C-terminal Cro protein chameleon sequences. / Anderson, William J.; Van Dorn, Laura O.; Ingram, Wendy M.; Cordes, Matthew Hj.

In: Protein Engineering, Design and Selection, Vol. 24, No. 9, 09.2011, p. 765-771.

Research output: Contribution to journalArticle

Anderson, William J. ; Van Dorn, Laura O. ; Ingram, Wendy M. ; Cordes, Matthew Hj. / Evolutionary bridges to new protein folds : Design of C-terminal Cro protein chameleon sequences. In: Protein Engineering, Design and Selection. 2011 ; Vol. 24, No. 9. pp. 765-771.
@article{e96b3fe2e13341dfaeaf766e0badfda0,
title = "Evolutionary bridges to new protein folds: Design of C-terminal Cro protein chameleon sequences",
abstract = "Regions of amino-acid sequence that are compatible with multiple folds may facilitate evolutionary transitions in protein structure. In a previous study, we described a heuristically designed chameleon sequence (SASF1, structurally ambivalent sequence fragment 1) that could adopt either of two naturally occurring conformations (α-helical or β-sheet) when incorporated as part of the C-terminal dimerization subdomain of two structurally divergent transcription factors, P22 Cro and λ Cro. Here we describe longer chameleon designs (SASF2 and SASF3) that in the case of SASF3 correspond to the full C-terminal half of the ordered region of a P22 Cro/λ Cro sequence alignment (residues 34-57). P22-SASF2 and λWDD-SASF2 show moderate thermal stability in denaturation curves monitored by circular dichroism (Tm values of 46 and 55°C, respectively), while P22-SASF3 and λWDD-SASF3 have somewhat reduced stability (Tm values of 33 and 49°C, respectively). 13C and 1H NMR secondary chemical shift analysis confirms two C-terminal α-helices for P22-SASF2 (residues 36-45 and 54-57) and two C-terminal β-strands for λWDD-SASF2 (residues 4045 and 5052), corresponding to secondary structure locations in the two parent sequences. Backbone relaxation data show that both chameleon sequences have a relatively well-ordered structure. Comparisons of 15N- 1H correlation spectra for SASF2 and SASF3-containing proteins strongly suggest that SASF3 retains the chameleonism of SASF2. Both Cro C-terminal conformations can be encoded in a single sequence, showing the plausibility of linking different Cro folds by smooth evolutionary transitions. The N-terminal subdomain, though largely conserved in structure, also exerts an important contextual influence on the structure of the C-terminal region.",
keywords = "chameleon sequence, NMR spectroscopy, protein design",
author = "Anderson, {William J.} and {Van Dorn}, {Laura O.} and Ingram, {Wendy M.} and Cordes, {Matthew Hj}",
year = "2011",
month = "9",
doi = "10.1093/protein/gzr027",
language = "English (US)",
volume = "24",
pages = "765--771",
journal = "Protein Engineering, Design and Selection",
issn = "1741-0126",
publisher = "Oxford University Press",
number = "9",

}

TY - JOUR

T1 - Evolutionary bridges to new protein folds

T2 - Design of C-terminal Cro protein chameleon sequences

AU - Anderson, William J.

AU - Van Dorn, Laura O.

AU - Ingram, Wendy M.

AU - Cordes, Matthew Hj

PY - 2011/9

Y1 - 2011/9

N2 - Regions of amino-acid sequence that are compatible with multiple folds may facilitate evolutionary transitions in protein structure. In a previous study, we described a heuristically designed chameleon sequence (SASF1, structurally ambivalent sequence fragment 1) that could adopt either of two naturally occurring conformations (α-helical or β-sheet) when incorporated as part of the C-terminal dimerization subdomain of two structurally divergent transcription factors, P22 Cro and λ Cro. Here we describe longer chameleon designs (SASF2 and SASF3) that in the case of SASF3 correspond to the full C-terminal half of the ordered region of a P22 Cro/λ Cro sequence alignment (residues 34-57). P22-SASF2 and λWDD-SASF2 show moderate thermal stability in denaturation curves monitored by circular dichroism (Tm values of 46 and 55°C, respectively), while P22-SASF3 and λWDD-SASF3 have somewhat reduced stability (Tm values of 33 and 49°C, respectively). 13C and 1H NMR secondary chemical shift analysis confirms two C-terminal α-helices for P22-SASF2 (residues 36-45 and 54-57) and two C-terminal β-strands for λWDD-SASF2 (residues 4045 and 5052), corresponding to secondary structure locations in the two parent sequences. Backbone relaxation data show that both chameleon sequences have a relatively well-ordered structure. Comparisons of 15N- 1H correlation spectra for SASF2 and SASF3-containing proteins strongly suggest that SASF3 retains the chameleonism of SASF2. Both Cro C-terminal conformations can be encoded in a single sequence, showing the plausibility of linking different Cro folds by smooth evolutionary transitions. The N-terminal subdomain, though largely conserved in structure, also exerts an important contextual influence on the structure of the C-terminal region.

AB - Regions of amino-acid sequence that are compatible with multiple folds may facilitate evolutionary transitions in protein structure. In a previous study, we described a heuristically designed chameleon sequence (SASF1, structurally ambivalent sequence fragment 1) that could adopt either of two naturally occurring conformations (α-helical or β-sheet) when incorporated as part of the C-terminal dimerization subdomain of two structurally divergent transcription factors, P22 Cro and λ Cro. Here we describe longer chameleon designs (SASF2 and SASF3) that in the case of SASF3 correspond to the full C-terminal half of the ordered region of a P22 Cro/λ Cro sequence alignment (residues 34-57). P22-SASF2 and λWDD-SASF2 show moderate thermal stability in denaturation curves monitored by circular dichroism (Tm values of 46 and 55°C, respectively), while P22-SASF3 and λWDD-SASF3 have somewhat reduced stability (Tm values of 33 and 49°C, respectively). 13C and 1H NMR secondary chemical shift analysis confirms two C-terminal α-helices for P22-SASF2 (residues 36-45 and 54-57) and two C-terminal β-strands for λWDD-SASF2 (residues 4045 and 5052), corresponding to secondary structure locations in the two parent sequences. Backbone relaxation data show that both chameleon sequences have a relatively well-ordered structure. Comparisons of 15N- 1H correlation spectra for SASF2 and SASF3-containing proteins strongly suggest that SASF3 retains the chameleonism of SASF2. Both Cro C-terminal conformations can be encoded in a single sequence, showing the plausibility of linking different Cro folds by smooth evolutionary transitions. The N-terminal subdomain, though largely conserved in structure, also exerts an important contextual influence on the structure of the C-terminal region.

KW - chameleon sequence

KW - NMR spectroscopy

KW - protein design

UR - http://www.scopus.com/inward/record.url?scp=80052326726&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=80052326726&partnerID=8YFLogxK

U2 - 10.1093/protein/gzr027

DO - 10.1093/protein/gzr027

M3 - Article

C2 - 21676898

AN - SCOPUS:80052326726

VL - 24

SP - 765

EP - 771

JO - Protein Engineering, Design and Selection

JF - Protein Engineering, Design and Selection

SN - 1741-0126

IS - 9

ER -