Details
| Originalsprache | Englisch |
|---|---|
| Seiten (von - bis) | 59-70 |
| Seitenumfang | 12 |
| Fachzeitschrift | Journal of Bioenergetics and Biomembranes |
| Jahrgang | 45 |
| Ausgabenummer | 1-2 |
| Publikationsstatus | Veröffentlicht - 14 Okt. 2012 |
Abstract
The C-terminus (CT) of rCx46 consists of 186 residues (H230-I416). Recent studies showed that rCx4628.2, truncated after H243, altered the formation of functional hemichannels when expressed in Xenopus oocytes, while rCx4637.7, truncated after A333 formed gap junction hemichannels similarly to rCx46wt. To analyze the role of the CT up to A333 in functional expression with cell imaging and dye-transfer techniques, different mutants were generated by C-terminal truncation between H243-A333, labeled with EGFP and expressed in HeLa cells. These rCx46 variants were characterized according to their compartmentalization in organelles, their presence in microscopic detectable vesicles and their ability to form gap junction plaques. rCx46 truncated after A311 (rCx4635.3) was compartmentalized, was found in vesicles and formed functional gap junction plaques similarly to rCx46wt. With a truncation after P284 (rCx4632.6), the protein was not compartmentalized and the amount of vesicles containing the protein were reduced; however, functional gap junction plaque formation was not affected as compared to rCx4635.3. rCx4628.2 did not form functional gap junction plaques; it was not found in vesicles or in cellular compartments. Live-cell imaging and detection of annular junctions for rCx4632.6 and rCx4635.3 revealed that the truncation after P284 reduced the frequency of vesicle budding from gap junction plaques and the formation of annular junctions. These results suggest that the C-terminal region of rCx46 up to A311 (rCx4635.3) is necessary for its correct compartmentalization and internalization in the form of annular junctions, while the H230-P284 C-terminal region (rCx4632.6) is sufficient for the formation of dye coupled gap junction channels.
ASJC Scopus Sachgebiete
- Biochemie, Genetik und Molekularbiologie (insg.)
- Physiologie
- Biochemie, Genetik und Molekularbiologie (insg.)
- Zellbiologie
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in: Journal of Bioenergetics and Biomembranes, Jahrgang 45, Nr. 1-2, 14.10.2012, S. 59-70.
Publikation: Beitrag in Fachzeitschrift › Artikel › Forschung › Peer-Review
}
TY - JOUR
T1 - The role of the C-terminus in functional expression and internalization of rat connexin46 (rCx46)
AU - Schlingmann, Barbara
AU - Schadzek, Patrik
AU - Hemmerling, Franziska
AU - Schaarschmidt, Frank
AU - Heisterkamp, Alexander
AU - Ngezahayo, Anaclet
PY - 2012/10/14
Y1 - 2012/10/14
N2 - The C-terminus (CT) of rCx46 consists of 186 residues (H230-I416). Recent studies showed that rCx4628.2, truncated after H243, altered the formation of functional hemichannels when expressed in Xenopus oocytes, while rCx4637.7, truncated after A333 formed gap junction hemichannels similarly to rCx46wt. To analyze the role of the CT up to A333 in functional expression with cell imaging and dye-transfer techniques, different mutants were generated by C-terminal truncation between H243-A333, labeled with EGFP and expressed in HeLa cells. These rCx46 variants were characterized according to their compartmentalization in organelles, their presence in microscopic detectable vesicles and their ability to form gap junction plaques. rCx46 truncated after A311 (rCx4635.3) was compartmentalized, was found in vesicles and formed functional gap junction plaques similarly to rCx46wt. With a truncation after P284 (rCx4632.6), the protein was not compartmentalized and the amount of vesicles containing the protein were reduced; however, functional gap junction plaque formation was not affected as compared to rCx4635.3. rCx4628.2 did not form functional gap junction plaques; it was not found in vesicles or in cellular compartments. Live-cell imaging and detection of annular junctions for rCx4632.6 and rCx4635.3 revealed that the truncation after P284 reduced the frequency of vesicle budding from gap junction plaques and the formation of annular junctions. These results suggest that the C-terminal region of rCx46 up to A311 (rCx4635.3) is necessary for its correct compartmentalization and internalization in the form of annular junctions, while the H230-P284 C-terminal region (rCx4632.6) is sufficient for the formation of dye coupled gap junction channels.
AB - The C-terminus (CT) of rCx46 consists of 186 residues (H230-I416). Recent studies showed that rCx4628.2, truncated after H243, altered the formation of functional hemichannels when expressed in Xenopus oocytes, while rCx4637.7, truncated after A333 formed gap junction hemichannels similarly to rCx46wt. To analyze the role of the CT up to A333 in functional expression with cell imaging and dye-transfer techniques, different mutants were generated by C-terminal truncation between H243-A333, labeled with EGFP and expressed in HeLa cells. These rCx46 variants were characterized according to their compartmentalization in organelles, their presence in microscopic detectable vesicles and their ability to form gap junction plaques. rCx46 truncated after A311 (rCx4635.3) was compartmentalized, was found in vesicles and formed functional gap junction plaques similarly to rCx46wt. With a truncation after P284 (rCx4632.6), the protein was not compartmentalized and the amount of vesicles containing the protein were reduced; however, functional gap junction plaque formation was not affected as compared to rCx4635.3. rCx4628.2 did not form functional gap junction plaques; it was not found in vesicles or in cellular compartments. Live-cell imaging and detection of annular junctions for rCx4632.6 and rCx4635.3 revealed that the truncation after P284 reduced the frequency of vesicle budding from gap junction plaques and the formation of annular junctions. These results suggest that the C-terminal region of rCx46 up to A311 (rCx4635.3) is necessary for its correct compartmentalization and internalization in the form of annular junctions, while the H230-P284 C-terminal region (rCx4632.6) is sufficient for the formation of dye coupled gap junction channels.
KW - Annular junctions
KW - C-terminus
KW - Connexin internalization
KW - rCx46
KW - Trafficking
KW - Vesicle budding
UR - http://www.scopus.com/inward/record.url?scp=84874114283&partnerID=8YFLogxK
U2 - 10.1007/s10863-012-9480-x
DO - 10.1007/s10863-012-9480-x
M3 - Article
C2 - 23065326
AN - SCOPUS:84874114283
VL - 45
SP - 59
EP - 70
JO - Journal of Bioenergetics and Biomembranes
JF - Journal of Bioenergetics and Biomembranes
SN - 0145-479X
IS - 1-2
ER -