PMID- 8401230
OWN - NLM
STAT- MEDLINE
DCOM- 19931123
LR  - 20181113
IS  - 0961-8368 (Print)
IS  - 1469-896X (Electronic)
IS  - 0961-8368 (Linking)
VI  - 2
IP  - 9
DP  - 1993 Sep
TI  - Bacterial expression and characterization of the CREB bZip module: circular 
      dichroism and 2D 1H-NMR studies.
PG  - 1461-71
AB  - In this paper we describe the expression and purification from bacteria of the 
      recombinant basic leucine zipper (bZip) domain of the cAMP response element 
      binding protein, CREB327. The bZip peptide, CREB259-327, purified to near 
      homogeneity, maintains the sequence-specific CRE site recognition demonstrated by 
      in vitro competition assays. Alkylation of the three cysteine residues of 
      CREB259-327 was employed to prevent aggregation of the peptide due to cysteine 
      oxidation. The Kd of the purified native and modified CREB259-327 for the CRE 
      site was determined by gel retardation assays to be on the order of 10(-7) M. We 
      employed CD spectroscopy to study the folding properties of the native and 
      modified CREB259-327. The CD analyses of the native/modified CREB259-327 peptide 
      demonstrated a 20% increase in the alpha-helical content upon binding to the cAMP 
      response-element. Only a 5% increase in the alpha-helical content of CREB259-327 
      is observed upon binding to the AP-1 site. This observation contrasts with CREB 
      from the GCN4 protein (Weiss, M.A., et al., 1990, Nature 347, 575-578). In 
      addition, the two-dimensional (2D) 1H-NMR studies of the bZip CREB peptide 
      further support the distinct features of the CREB protein, in comparison to GCN4. 
      Analysis by CD and 2D NMR of the dimerization domain of CREB suggests that the 
      distinct DNA binding characteristics of CREB reside in the basic portion of the 
      bZip module.
FAU - Santiago-Rivera, Z I
AU  - Santiago-Rivera ZI
AD  - Department of Chemistry, School of Science, Purdue University, West Lafayette, 
      Indiana 47907.
FAU - Williams, J S
AU  - Williams JS
FAU - Gorenstein, D G
AU  - Gorenstein DG
FAU - Andrisani, O M
AU  - Andrisani OM
LA  - eng
GR  - AI27744/AI/NIAID NIH HHS/United States
GR  - AI727713/AI/NIAID NIH HHS/United States
GR  - R29 DK44533/DK/NIDDK NIH HHS/United States
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, Non-P.H.S.
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - Protein Sci
JT  - Protein science : a publication of the Protein Society
JID - 9211750
RN  - 0 (Cyclic AMP Response Element-Binding Protein)
RN  - 0 (Peptide Fragments)
RN  - 9007-49-2 (DNA)
RN  - K848JZ4886 (Cysteine)
SB  - IM
MH  - Amino Acid Sequence
MH  - Base Sequence
MH  - Chromatography, High Pressure Liquid
MH  - *Circular Dichroism
MH  - Cyclic AMP Response Element-Binding Protein/*chemistry/*genetics/metabolism
MH  - Cysteine/metabolism
MH  - DNA/metabolism
MH  - Electrophoresis, Polyacrylamide Gel
MH  - Escherichia coli/*genetics
MH  - *Gene Expression
MH  - *Leucine Zippers
MH  - *Magnetic Resonance Spectroscopy
MH  - Methylation
MH  - Molecular Sequence Data
MH  - Peptide Fragments/chemistry/genetics/metabolism
MH  - Protein Conformation
MH  - Protein Structure, Secondary
MH  - Transformation, Bacterial
PMC - PMC2142467
EDAT- 1993/09/01 00:00
MHDA- 1993/09/01 00:01
PMCR- 1994/03/01
CRDT- 1993/09/01 00:00
PHST- 1993/09/01 00:00 [pubmed]
PHST- 1993/09/01 00:01 [medline]
PHST- 1993/09/01 00:00 [entrez]
PHST- 1994/03/01 00:00 [pmc-release]
AID - 10.1002/pro.5560020910 [doi]
PST - ppublish
SO  - Protein Sci. 1993 Sep;2(9):1461-71. doi: 10.1002/pro.5560020910.