Linked Life Data

7592746

Source:http://linkedlifedata.com/resource/pubmed/id/7592746

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
43
pubmed:dateCreated
1995-12-14
pubmed:abstractText
The membrane topology of the rat endoplasmic reticulum (ER) and sarcoplasmic reticulum (SR) Ca2+ ATPases were investigated using in vitro transcription/translation of fusion vectors containing DNA sequences encoding putative membrane-spanning domains. The sequences of these Ca2+ ATPases are identical except for the COOH-terminal end, which contains an additional predicted transmembrane segment in the ER ATPase. The M0 and M1 fusion vectors (Bamberg, K., and Sachs, G. (1994) J. Biol. Chem. 269, 16909-16919) encode the NH2-terminal 101 (M0 vector) or 139 (M1 vector) amino acids of the H,K-ATPase alpha subunit followed by a linker region for insertion of putative transmembrane sequences and, finally, the COOH-terminal 177 amino acids of the H,K-ATPase beta subunit containing five N-linked glycosylation consensus sequences. The linker region was replaced by the putative transmembrane domains of the Ca2+ ATPases, either individually or in pairs. Transcription and translation were performed using [35S]methionine in a reticulocyte lysate system in the absence or presence of canine pancreatic microsomes. The translated fusion protein was identified by autoradiography following separation using SDS-polyacrylamide gel electrophoresis. When testing single transmembrane segments, this method detects signal anchor activity with M0 or stop transfer activity with M1. The first four predicted SERCA transmembrane domains acted as both signal anchor and stop transfer sequences. A construct containing the fifth predicted transmembrane segment was able to act only as a stop transfer sequence. The sixth transmembrane segment did not insert cotranslationally into the membrane. The seventh was able to act as both a signal anchor and stop transfer sequence, and the eighth showed stop transfer ability in the M1 vector. The ninth transmembrane segment had both signal anchor and stop transfer capacity, whereas the tenth transmembrane segment showed only stop transfer sequence properties. The eleventh transmembrane sequence, unique to the ER Ca2+ ATPase, had both signal anchor and stop transfer properties. These translation data provide direct experimental evidence for 8 or 9 of the 10 or 11 predicted transmembrane sequences in the current topological models for the SR or ER Ca2+ ATPases, respectively.
pubmed:grant
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Oct
pubmed:issn
0021-9258
pubmed:author
pubmed:issnType
Print
pubmed:day
27
pubmed:volume
270
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
25678-84
pubmed:dateRevised
2010-11-18
pubmed:meshHeading
pubmed:year
1995
pubmed:articleTitle
The membrane topology of the rat sarcoplasmic and endoplasmic reticulum calcium ATPases by in vitro translation scanning.
pubmed:affiliation
Department of Medicine and Physiology, UCLA 90073, USA.
pubmed:publicationType
Journal Article, Comparative Study, Research Support, U.S. Gov't, P.H.S., Research Support, U.S. Gov't, Non-P.H.S.