PMID- 27586331
OWN - NLM
STAT- MEDLINE
DCOM- 20170807
LR  - 20181113
IS  - 1557-7988 (Electronic)
IS  - 0076-6879 (Print)
IS  - 0076-6879 (Linking)
VI  - 580
DP  - 2016
TI  - Methods for Solving Highly Symmetric De Novo Designed Metalloproteins: 
      Crystallographic Examination of a Novel Three-Stranded Coiled-Coil Structure 
      Containing d-Amino Acids.
PG  - 135-48
LID - S0076-6879(16)30042-8 [pii]
LID - 10.1016/bs.mie.2016.05.007 [doi]
AB  - The core objective of de novo metalloprotein design is to define metal-protein 
      relationships that control the structure and function of metal centers by using 
      simplified proteins. An essential requirement to achieve this goal is to obtain 
      high resolution structural data using either NMR or crystallographic studies in 
      order to evaluate successful design. X-ray crystal structures have proven that a 
      four heptad repeat scaffold contained in the three-stranded coiled coil (3SCC), 
      called CoilSer (CS), provides an excellent motif for modeling a three Cys binding 
      environment capable of chelating metals into geometries that resemble heavy metal 
      sites in metalloregulatory systems. However, new generations of more complicated 
      designs that feature, for example, a d-amino acid or multiple metal ligand sites 
      in the helical sequence require a more stable construct. In doing so, an extra 
      heptad was introduced into the original CS sequence, yielding a GRAND-CoilSer 
      (GRAND-CS) to retain the 3SCC folding. An apo-(GRAND-CSL12DLL16C)3 crystal 
      structure, designed for Cd(II)S3 complexation, proved to be a well-folded 
      parallel 3SCC. Because this structure is novel, protocols for crystallization, 
      structural determination, and refinements of the apo-(GRAND-CSL12DLL16C)3 are 
      described. This report should be generally useful for future crystallographic 
      studies of related coiled-coil designs.
CI  - (c) 2016 Elsevier Inc. All rights reserved.
FAU - Ruckthong, L
AU  - Ruckthong L
AD  - Department of Chemistry, University of Michigan, Ann Arbor, MI, United States.
FAU - Stuckey, J A
AU  - Stuckey JA
AD  - Life Sciences Institute, University of Michigan, Ann Arbor, MI, United States.
FAU - Pecoraro, V L
AU  - Pecoraro VL
AD  - Department of Chemistry, University of Michigan, Ann Arbor, MI, United States. 
      Electronic address: vlpec@umich.edu.
LA  - eng
GR  - R01 ES012236/ES/NIEHS NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, Non-P.H.S.
DEP - 20160623
PL  - United States
TA  - Methods Enzymol
JT  - Methods in enzymology
JID - 0212271
RN  - 0 (Amino Acids)
RN  - 0 (Ligands)
RN  - 0 (Metalloproteins)
RN  - 0 (Metals)
RN  - 0 (Peptides)
SB  - IM
MH  - Amino Acid Sequence/genetics
MH  - Amino Acids/*chemistry/genetics
MH  - Binding Sites
MH  - Crystallography, X-Ray
MH  - Ligands
MH  - Metalloproteins/chemical synthesis/*chemistry/genetics
MH  - Metals
MH  - Models, Molecular
MH  - Peptides/*chemistry
MH  - *Protein Structure, Secondary
PMC - PMC5237576
MID - NIHMS841510
OTO - NOTNLM
OT  - De novo protein design
OT  - Three-stranded coiled coil
OT  - X-ray crystallography
OT  - d-Amino acid
EDAT- 2016/09/03 06:00
MHDA- 2017/08/08 06:00
PMCR- 2017/01/15
CRDT- 2016/09/03 06:00
PHST- 2016/09/03 06:00 [entrez]
PHST- 2016/09/03 06:00 [pubmed]
PHST- 2017/08/08 06:00 [medline]
PHST- 2017/01/15 00:00 [pmc-release]
AID - S0076-6879(16)30042-8 [pii]
AID - 10.1016/bs.mie.2016.05.007 [doi]
PST - ppublish
SO  - Methods Enzymol. 2016;580:135-48. doi: 10.1016/bs.mie.2016.05.007. Epub 2016 Jun 
      23.