PMID- 10097073
OWN - NLM
STAT- MEDLINE
DCOM- 19990512
LR  - 20190501
IS  - 0027-8424 (Print)
IS  - 0027-8424 (Linking)
VI  - 96
IP  - 7
DP  - 1999 Mar 30
TI  - Biochemical characterization of Wnt-frizzled interactions using a soluble,
      biologically active vertebrate Wnt protein.
PG  - 3546-51
AB  - Biochemical studies of Wnt signaling have been hampered by difficulties in
      obtaining large quantities of soluble, biologically active Wnt proteins. In this 
      paper, we report the production in Drosophila S2 cells of biologically active
      Xenopus Wnt8 (XWnt8). Epitope- or alkaline phosphatase-tagged XWnt8 proteins are 
      secreted by concentrated S2 cells in a form that is suitable for quantitative
      biochemical experiments with yields of 5 and 0.5 mg per liter, respectively.
      Conditions also are described for the production in 293 cells of an IgG fusion of
      the cysteine-rich domain (CRD) of mouse Frizzled 8 with a yield of 20 mg/liter.
      We demonstrate the use of these proteins for studying the interactions between
      soluble XWnt8 and various Frizzled proteins, membrane anchored or secreted CRDs, 
      and a set of insertion mutants in the CRD of Drosophila Frizzled 2. In a solid
      phase binding assay, the affinity of the XWnt8-alkaline phosphatase fusion for
      the purified mouse Frizzled 8-CRD-IgG fusion is approximately 9 nM.
FAU - Hsieh, J C
AU  - Hsieh JC
AD  - Department of Molecular Biology and Genetics, Johns Hopkins University School of 
      Medicine, Baltimore, MD 21205, USA.
FAU - Rattner, A
AU  - Rattner A
FAU - Smallwood, P M
AU  - Smallwood PM
FAU - Nathans, J
AU  - Nathans J
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Proc Natl Acad Sci U S A
JT  - Proceedings of the National Academy of Sciences of the United States of America
JID - 7505876
RN  - 0 (Cytoskeletal Proteins)
RN  - 0 (Drosophila Proteins)
RN  - 0 (Frizzled Receptors)
RN  - 0 (Proteins)
RN  - 0 (Receptors, G-Protein-Coupled)
RN  - 0 (Receptors, Neurotransmitter)
RN  - 0 (Recombinant Proteins)
RN  - 0 (Wnt Proteins)
RN  - 0 (Zebrafish Proteins)
RN  - 0 (fz2 protein, Drosophila)
RN  - 0 (wnt8a protein, zebrafish)
SB  - IM
MH  - Animals
MH  - Base Sequence
MH  - COS Cells
MH  - Cell Line
MH  - Cytoskeletal Proteins
MH  - *Drosophila Proteins
MH  - Drosophila melanogaster
MH  - Frizzled Receptors
MH  - Humans
MH  - Kinetics
MH  - Mice
MH  - Molecular Sequence Data
MH  - Protein Structure, Secondary
MH  - Proteins/chemistry/genetics/*metabolism
MH  - Receptors, G-Protein-Coupled
MH  - Receptors, Neurotransmitter/chemistry/genetics/*metabolism
MH  - Recombinant Proteins/biosynthesis/chemistry/metabolism
MH  - Sequence Alignment
MH  - Sequence Homology, Amino Acid
MH  - Transfection
MH  - Vertebrates
MH  - Wnt Proteins
MH  - Xenopus
MH  - Zebrafish Proteins
PMC - PMC22330
EDAT- 1999/03/31 00:00
MHDA- 1999/03/31 00:01
CRDT- 1999/03/31 00:00
PHST- 1999/03/31 00:00 [pubmed]
PHST- 1999/03/31 00:01 [medline]
PHST- 1999/03/31 00:00 [entrez]
AID - 10.1073/pnas.96.7.3546 [doi]
PST - ppublish
SO  - Proc Natl Acad Sci U S A. 1999 Mar 30;96(7):3546-51. doi: 10.1073/pnas.96.7.3546.