PMID- 10072769
OWN - NLM
STAT- MEDLINE
DCOM- 19990413
LR  - 20190707
IS  - 0378-1119 (Print)
IS  - 0378-1119 (Linking)
VI  - 228
IP  - 1-2
DP  - 1999 Mar 4
TI  - Selection of cDNAs encoding putative type II membrane proteins on the cell
      surface from a human full-length cDNA bank.
PG  - 161-7
AB  - We have developed a simple method to test whether a hydrophobic segment near the 
      N-terminus of a protein functions as a type II signal anchor (SA) in which the
      N-terminus faces the cytoplasm. A cDNA fragment containing the putative SA
      sequence of a target clone was fused in-frame to the 5' end of a cDNA fragment
      encoding the protease domain of urokinase-type plasminogen activator (u-PA). The 
      resulting fused gene was expressed in COS7 cells. Fibrinolytic activity on the
      cell surface was measured by placing a fibrin sheet in contact with the
      transfected COS7 cells after removing the medium. When the cDNA fragment encoded 
      a SA, the fibrin sheet was lysed by the u-PA expressed on the cell surface. The
      fibrinolytic activity was not detected in the culture medium, suggesting that the
      u-PA remains on the cell surface anchored via the SA in the membrane without
      being cleaved by signal peptidase. This fibrin sheet method was successfully
      applied to select five novel cDNA clones encoding putative type II membrane
      proteins from a human full-length cDNA bank.
FAU - Yokoyama-Kobayashi, M
AU  - Yokoyama-Kobayashi M
AD  - Kato Cytoprotein Network Project, ERATO, JST, 4-4-1 Nishi-Ohnuma, Sagamihara,
      Kanagawa 229-0012, Japan.
FAU - Yamaguchi, T
AU  - Yamaguchi T
FAU - Sekine, S
AU  - Sekine S
FAU - Kato, S
AU  - Kato S
LA  - eng
SI  - GENBANK/AB015628
SI  - GENBANK/AB015629
SI  - GENBANK/AB015630
SI  - GENBANK/AB015631
SI  - GENBANK/AB015632
SI  - GENBANK/AB015633
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - Netherlands
TA  - Gene
JT  - Gene
JID - 7706761
RN  - 0 (DNA, Complementary)
RN  - 0 (Membrane Proteins)
RN  - 0 (Protein Sorting Signals)
RN  - 0 (Recombinant Fusion Proteins)
RN  - EC 3.4.- (Endopeptidases)
RN  - EC 3.4.21.73 (Urokinase-Type Plasminogen Activator)
SB  - IM
MH  - Animals
MH  - Binding Sites
MH  - COS Cells
MH  - Cell Membrane/metabolism
MH  - DNA, Complementary/chemistry/*genetics
MH  - Endopeptidases/genetics/metabolism
MH  - Fibrinolysis
MH  - *Gene Library
MH  - Humans
MH  - Membrane Proteins/*genetics
MH  - Molecular Sequence Data
MH  - Protein Sorting Signals/genetics
MH  - Recombinant Fusion Proteins/genetics/metabolism
MH  - U937 Cells
MH  - Urokinase-Type Plasminogen Activator/genetics/metabolism
EDAT- 1999/03/12 00:00
MHDA- 1999/03/12 00:01
CRDT- 1999/03/12 00:00
PHST- 1999/03/12 00:00 [pubmed]
PHST- 1999/03/12 00:01 [medline]
PHST- 1999/03/12 00:00 [entrez]
AID - S0378-1119(99)00004-9 [pii]
AID - 10.1016/s0378-1119(99)00004-9 [doi]
PST - ppublish
SO  - Gene. 1999 Mar 4;228(1-2):161-7. doi: 10.1016/s0378-1119(99)00004-9.