SNAP-23 in rat kidney : colocalization with aquaporin-2 in collecting duct vesicles
Vesicle targeting proteins ("SNAREs") have been proposed to direct vasopressin-induced trafficking of aquaporin-2 water channels in kidney collecting ducts. A newly identified SNARE protein, SNAP-23, is proposed to mediate vesicle targeting to the plasma membrane in diverse tissues. The current studies were done to determine whether SNAP-23 is expressed in collecting ducts with an intracellular distribution compatible with a role in aquaporin-2 trafficking. RT-PCR demonstrated SNAP-23 mRNA in microdissected collecting ducts and other tubular segments including the proximal tubule and thick ascending limb. Immunoblotting using a polyclonal antibody raised against a COOH-terminal peptide revealed a solitary band at an apparent molecular mass of 30 kDa in renal medullary membrane fractions and inner medullary collecting duct suspensions. Differential centrifugation revealed that SNAP-23 is present in membrane fractions including the low-density fraction enriched in intracellular vesicles. Immunocytochemistry revealed SNAP-23 labeling at both the apex and the cytoplasm of collecting duct principal cells. Immunoblotting of intracellular vesicles immunoisolated using an aquaporin-2 antibody revealed the presence of both SNAP-23 and synaptobrevin-2 (VAMP-2) in aquaporin-2-bearing vesicles. We conclude that SNAP-23 is strongly expressed in collecting duct principal cells, consistent with a role in vasopressin-regulated trafficking of aquaporin-2. However, localization of SNAP-23 in both intracytoplasmic vesicles and plasma membranes suggests a function different from that originally proposed for SNAP-25 in synaptic vesicle targeting.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
1998 |
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| Erschienen: |
1998 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:275 |
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| Enthalten in: |
The American journal of physiology - 275(1998), 5 vom: 22. Nov., Seite F752-60 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Inoue, T [VerfasserIn] |
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| Links: |
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| Anmerkungen: |
Date Completed 10.12.1998 Date Revised 22.02.2019 published: Print Citation Status MEDLINE |
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| doi: |
10.1152/ajprenal.1998.275.5.F752 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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| PPN (Katalog-ID): |
NLM097530018 |
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| 100 | 1 | |a Inoue, T |e verfasserin |4 aut | |
| 245 | 1 | 0 | |a SNAP-23 in rat kidney |b colocalization with aquaporin-2 in collecting duct vesicles |
| 264 | 1 | |c 1998 | |
| 336 | |a Text |b txt |2 rdacontent | ||
| 337 | |a ƒaComputermedien |b c |2 rdamedia | ||
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| 500 | |a Date Completed 10.12.1998 | ||
| 500 | |a Date Revised 22.02.2019 | ||
| 500 | |a published: Print | ||
| 500 | |a Citation Status MEDLINE | ||
| 520 | |a Vesicle targeting proteins ("SNAREs") have been proposed to direct vasopressin-induced trafficking of aquaporin-2 water channels in kidney collecting ducts. A newly identified SNARE protein, SNAP-23, is proposed to mediate vesicle targeting to the plasma membrane in diverse tissues. The current studies were done to determine whether SNAP-23 is expressed in collecting ducts with an intracellular distribution compatible with a role in aquaporin-2 trafficking. RT-PCR demonstrated SNAP-23 mRNA in microdissected collecting ducts and other tubular segments including the proximal tubule and thick ascending limb. Immunoblotting using a polyclonal antibody raised against a COOH-terminal peptide revealed a solitary band at an apparent molecular mass of 30 kDa in renal medullary membrane fractions and inner medullary collecting duct suspensions. Differential centrifugation revealed that SNAP-23 is present in membrane fractions including the low-density fraction enriched in intracellular vesicles. Immunocytochemistry revealed SNAP-23 labeling at both the apex and the cytoplasm of collecting duct principal cells. Immunoblotting of intracellular vesicles immunoisolated using an aquaporin-2 antibody revealed the presence of both SNAP-23 and synaptobrevin-2 (VAMP-2) in aquaporin-2-bearing vesicles. We conclude that SNAP-23 is strongly expressed in collecting duct principal cells, consistent with a role in vasopressin-regulated trafficking of aquaporin-2. However, localization of SNAP-23 in both intracytoplasmic vesicles and plasma membranes suggests a function different from that originally proposed for SNAP-25 in synaptic vesicle targeting | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a Research Support, Non-U.S. Gov't | |
| 650 | 4 | |a Research Support, U.S. Gov't, P.H.S. | |
| 650 | 7 | |a Aqp2 protein, rat |2 NLM | |
| 650 | 7 | |a Aquaporin 2 |2 NLM | |
| 650 | 7 | |a Aquaporin 6 |2 NLM | |
| 650 | 7 | |a Aquaporins |2 NLM | |
| 650 | 7 | |a Carrier Proteins |2 NLM | |
| 650 | 7 | |a Membrane Proteins |2 NLM | |
| 650 | 7 | |a Qb-SNARE Proteins |2 NLM | |
| 650 | 7 | |a Qc-SNARE Proteins |2 NLM | |
| 650 | 7 | |a R-SNARE Proteins |2 NLM | |
| 650 | 7 | |a RNA, Messenger |2 NLM | |
| 650 | 7 | |a SNAP23 protein, human |2 NLM | |
| 700 | 1 | |a Nielsen, S |e verfasserin |4 aut | |
| 700 | 1 | |a Mandon, B |e verfasserin |4 aut | |
| 700 | 1 | |a Terris, J |e verfasserin |4 aut | |
| 700 | 1 | |a Kishore, B K |e verfasserin |4 aut | |
| 700 | 1 | |a Knepper, M A |e verfasserin |4 aut | |
| 773 | 0 | 8 | |i Enthalten in |t The American journal of physiology |d 1945 |g 275(1998), 5 vom: 22. Nov., Seite F752-60 |w (DE-627)NLM000012386 |x 0002-9513 |7 nnns |
| 773 | 1 | 8 | |g volume:275 |g year:1998 |g number:5 |g day:22 |g month:11 |g pages:F752-60 |
| 856 | 4 | 0 | |u http://dx.doi.org/10.1152/ajprenal.1998.275.5.F752 |3 Volltext |
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