Abstract
Signal recognition particle (SRP) plays a critical role in the targeting of secretory proteins to cellular membranes. An essential component of SRP is the protein SRP54, which interacts not only with the nascent signal peptide, but also with the SRP RNA. To understand better how protein targeting occurs in the human system, the human SRP54 gene was cloned, sequenced, and the protein was expressed in bacteria and insect cells. Recombinant SRP54 was purified from both sources. The protein bound to SRP RNA in the presence of protein SRP19, and associated with the signal peptide of in vitro translated pre-prolactin. Comparative sequence analysis of human SRP54 with homologs from all three phylogenetic domains was combined with high-stringency protein secondary structure prediction. A conserved RNA-binding loop was predicted in the largely helical M-domain of SRP54. Contrary to general belief, the unusually high number of methionine residues clustered outside the predicted helices, thus indicating a mechanism of signal peptide recognition that may involve methionine-rich loops.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 197-207 |
| Number of pages | 11 |
| Journal | Gene |
| Volume | 207 |
| Issue number | 2 |
| DOIs | |
| State | Published - Jan 30 1998 |
| Externally published | Yes |
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Keywords
- Protein secretion
- Protein targeting
- RNA-protein interactions
- Signal peptide
ASJC Scopus subject areas
- Genetics
Cite this
Protein SRP54 of human signal recognition particle : Cloning, expression, and comparative analysis of functional sites. / Gowda, Krishne; Black, Shaun D.; Moeller, Ines; Sakakibara, Yoichi; Liu, Ming Cheh; Zwieb, Christian W.
In: Gene, Vol. 207, No. 2, 30.01.1998, p. 197-207.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Protein SRP54 of human signal recognition particle
T2 - Cloning, expression, and comparative analysis of functional sites
AU - Gowda, Krishne
AU - Black, Shaun D.
AU - Moeller, Ines
AU - Sakakibara, Yoichi
AU - Liu, Ming Cheh
AU - Zwieb, Christian W
PY - 1998/1/30
Y1 - 1998/1/30
N2 - Signal recognition particle (SRP) plays a critical role in the targeting of secretory proteins to cellular membranes. An essential component of SRP is the protein SRP54, which interacts not only with the nascent signal peptide, but also with the SRP RNA. To understand better how protein targeting occurs in the human system, the human SRP54 gene was cloned, sequenced, and the protein was expressed in bacteria and insect cells. Recombinant SRP54 was purified from both sources. The protein bound to SRP RNA in the presence of protein SRP19, and associated with the signal peptide of in vitro translated pre-prolactin. Comparative sequence analysis of human SRP54 with homologs from all three phylogenetic domains was combined with high-stringency protein secondary structure prediction. A conserved RNA-binding loop was predicted in the largely helical M-domain of SRP54. Contrary to general belief, the unusually high number of methionine residues clustered outside the predicted helices, thus indicating a mechanism of signal peptide recognition that may involve methionine-rich loops.
AB - Signal recognition particle (SRP) plays a critical role in the targeting of secretory proteins to cellular membranes. An essential component of SRP is the protein SRP54, which interacts not only with the nascent signal peptide, but also with the SRP RNA. To understand better how protein targeting occurs in the human system, the human SRP54 gene was cloned, sequenced, and the protein was expressed in bacteria and insect cells. Recombinant SRP54 was purified from both sources. The protein bound to SRP RNA in the presence of protein SRP19, and associated with the signal peptide of in vitro translated pre-prolactin. Comparative sequence analysis of human SRP54 with homologs from all three phylogenetic domains was combined with high-stringency protein secondary structure prediction. A conserved RNA-binding loop was predicted in the largely helical M-domain of SRP54. Contrary to general belief, the unusually high number of methionine residues clustered outside the predicted helices, thus indicating a mechanism of signal peptide recognition that may involve methionine-rich loops.
KW - Protein secretion
KW - Protein targeting
KW - RNA-protein interactions
KW - Signal peptide
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UR - http://www.scopus.com/inward/citedby.url?scp=0032579343&partnerID=8YFLogxK
U2 - 10.1016/S0378-1119(97)00627-6
DO - 10.1016/S0378-1119(97)00627-6
M3 - Article
C2 - 9511762
AN - SCOPUS:0032579343
VL - 207
SP - 197
EP - 207
JO - Gene
JF - Gene
SN - 0378-1119
IS - 2
ER -