PMID- 10199829
OWN - NLM
STAT- MEDLINE
DCOM- 19990518
LR  - 20190125
IS  - 0002-9513 (Print)
IS  - 0002-9513 (Linking)
VI  - 276
IP  - 4
DP  - 1999 Apr
TI  - Cloning of Trp1beta isoform from rat brain: immunodetection and localization of
      the endogenous Trp1 protein.
PG  - C969-79
LID - 10.1152/ajpcell.1999.276.4.C969 [doi]
AB  - The Trp gene product has been proposed as a candidate protein for the
      store-operated Ca2+ channel, but the Trp protein(s) has not been identified in
      any nonexcitable cell. We report here the cloning of a rat brain Trp1beta cDNA
      and detection and immunolocalization of the endogenous and expressed Trp1
      protein. A 400-bp product, with >95% homology to mouse Trp1, was amplified from
      rat submandibular gland RNA. Rat-specific primers were used for cloning of a
      full-length rat brain Trp1beta cDNA (rTrp1), encoding a protein of 759 amino
      acids. Northern blot analysis demonstrated the transcript in several rat and
      mouse tissues. The peptide (amino acids 523-536) was used to generate a
      polyclonal antiserum. The affinity-purified antibody 1) immunoprecipitated human 
      Trp1 (hTrp1) from transfected HEK-293 cells, 2) reacted with a protein of
      approximately 92 kDa, but not with hTrp3, in membranes of hTrp3-expressing
      HEK-293 cells, and 3) reacted with proteins of 92 and 56 kDa in human and rat
      brain membranes. Confocal microscopy and cell fractionation demonstrated that
      endogenous and expressed hTrp1 and expressed hTrp3 proteins were localized in the
      plasma membrane of HEK-293 cells, consistent with their proposed role in Ca2+
      influx. The data demonstrate for the first time the presence of Trp1 protein in a
      nonexcitable cell.
FAU - Wang, W
AU  - Wang W
AD  - Secretory Physiology Section, National Institute of Dental and Craniofacial
      Research, National Institutes of Health, Bethesda, Maryland 20892, USA.
FAU - O'Connell, B
AU  - O'Connell B
FAU - Dykeman, R
AU  - Dykeman R
FAU - Sakai, T
AU  - Sakai T
FAU - Delporte, C
AU  - Delporte C
FAU - Swaim, W
AU  - Swaim W
FAU - Zhu, X
AU  - Zhu X
FAU - Birnbaumer, L
AU  - Birnbaumer L
FAU - Ambudkar, I S
AU  - Ambudkar IS
LA  - eng
GR  - GM-54235/GM/NIGMS NIH HHS/United States
GR  - HL-45198/HL/NHLBI NIH HHS/United States
PT  - Journal Article
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - Am J Physiol
JT  - The American journal of physiology
JID - 0370511
RN  - 0 (Calcium Channels)
RN  - 0 (Protein Isoforms)
RN  - 0 (Recombinant Proteins)
RN  - 0 (TRPC Cation Channels)
RN  - 0 (transient receptor potential cation channel, subfamily C, member 1)
SB  - IM
MH  - Amino Acid Sequence
MH  - Animals
MH  - Brain/cytology/*metabolism
MH  - Calcium Channels/*analysis/chemistry/*genetics
MH  - Cattle
MH  - Cell Line
MH  - Cloning, Molecular
MH  - Humans
MH  - Mice
MH  - Molecular Sequence Data
MH  - Protein Isoforms/analysis/genetics
MH  - Rats
MH  - Recombinant Proteins/biosynthesis/chemistry
MH  - Sequence Alignment
MH  - Sequence Homology, Amino Acid
MH  - Submandibular Gland/metabolism
MH  - TRPC Cation Channels
MH  - Transcription, Genetic
MH  - Transfection
EDAT- 1999/04/13 00:00
MHDA- 1999/04/13 00:01
CRDT- 1999/04/13 00:00
PHST- 1999/04/13 00:00 [pubmed]
PHST- 1999/04/13 00:01 [medline]
PHST- 1999/04/13 00:00 [entrez]
AID - 10.1152/ajpcell.1999.276.4.C969 [doi]
PST - ppublish
SO  - Am J Physiol. 1999 Apr;276(4):C969-79. doi: 10.1152/ajpcell.1999.276.4.C969.