PMID- 10493790
OWN - NLM
STAT- MEDLINE
DCOM- 19991028
LR  - 20190613
IS  - 0006-2960 (Print)
IS  - 0006-2960 (Linking)
VI  - 38
IP  - 38
DP  - 1999 Sep 21
TI  - Expression, purification, and crystal structure determination of recombinant 
      human epidermal-type fatty acid binding protein.
PG  - 12229-39
AB  - We describe the crystal structure of human epidermal-type fatty acid binding 
      protein (E-FABP) that was recently found to be highly upregulated in human 
      psoriatic keratinocytes. To characterize E-FABP with respect to ligand-binding 
      properties and tertiary structure, we cloned the respective cDNA, overexpressed 
      the protein in Escherichia coli and purified it to homogeneity by a combination 
      of ion-exchange and size-exclusion chromatographic steps with a yield of 30 mg/L 
      broth. The purified protein revealed a 5-fold higher affinity for stearic acid 
      than for oleic and arachidonic acids. The crystal structure of recombinant human 
      E-FABP was determined to 2.05 A and refined to an R(factor) of 20.7%. The initial 
      residual electron density maps clearly showed the presence of a ligand, which was 
      identified as endogenous bacterial fatty acid. Within a central cavity of 252 
      A(3), this ligand is bound in a U-shaped conformation, its carboxyl group 
      interacting with tyrosine 131 and arginines 129 and 109, the latter via an 
      ordered water molecule. The E-FABP crystal structure is unique in the FABP family 
      because of the presence of a disulfide bridge between cysteines 120 and 127 that 
      may be physiologically as well as pathophysiologically relevant. Cysteines 67 and 
      87 are also in close vicinity but in contrast do not form a disulfide bridge. We 
      postulate that this protein belongs to a particular FABP subfamily whose members 
      share common structural as well as functional features.
FAU - Hohoff, C
AU  - Hohoff C
AD  - Institut fur Biochemie, Westfalische Wilhelms-Universitat Munster, Germany.
FAU - Borchers, T
AU  - Borchers T
FAU - Rustow, B
AU  - Rustow B
FAU - Spener, F
AU  - Spener F
FAU - van Tilbeurgh, H
AU  - van Tilbeurgh H
LA  - eng
SI  - PDB/1B56
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Biochemistry
JT  - Biochemistry
JID - 0370623
RN  - 0 (Carrier Proteins)
RN  - 0 (FABP5 protein, human)
RN  - 0 (FABP7 protein, human)
RN  - 0 (Fabp5 protein, mouse)
RN  - 0 (Fabp7 protein, mouse)
RN  - 0 (Fatty Acid-Binding Protein 7)
RN  - 0 (Fatty Acid-Binding Proteins)
RN  - 0 (Fatty Acids)
RN  - 0 (Myelin P2 Protein)
RN  - 0 (Neoplasm Proteins)
RN  - 0 (Nerve Tissue Proteins)
RN  - 0 (Recombinant Proteins)
RN  - 0 (Tumor Suppressor Proteins)
SB  - IM
MH  - Adipose Tissue
MH  - Amino Acid Sequence
MH  - Animals
MH  - Binding Sites/genetics
MH  - Carrier Proteins/*biosynthesis/*chemistry/genetics/metabolism
MH  - Cattle
MH  - Crystallization
MH  - Crystallography, X-Ray
MH  - Epidermis
MH  - Escherichia coli/genetics
MH  - Fatty Acid-Binding Protein 7
MH  - Fatty Acid-Binding Proteins
MH  - Fatty Acids/*metabolism
MH  - Humans
MH  - Mice
MH  - Models, Molecular
MH  - Molecular Sequence Data
MH  - Myelin P2 Protein/*biosynthesis/*chemistry/genetics/metabolism
MH  - *Neoplasm Proteins
MH  - *Nerve Tissue Proteins
MH  - Protein Structure, Tertiary
MH  - Recombinant Proteins/*biosynthesis/*chemistry/isolation & purification/metabolism
MH  - Skin
MH  - Structure-Activity Relationship
MH  - *Tumor Suppressor Proteins
EDAT- 1999/09/24 00:00
MHDA- 1999/09/24 00:01
CRDT- 1999/09/24 00:00
PHST- 1999/09/24 00:00 [pubmed]
PHST- 1999/09/24 00:01 [medline]
PHST- 1999/09/24 00:00 [entrez]
AID - bi990305u [pii]
AID - 10.1021/bi990305u [doi]
PST - ppublish
SO  - Biochemistry. 1999 Sep 21;38(38):12229-39. doi: 10.1021/bi990305u.