Expression of IGF receptors on alveolar macrophages: IGF-induced changes in InsPi formation, [Ca2+]i, and pHi
Expression of insulin-like growth factor-I (IGF-I) receptors and insulin-like growth factor-II/mannose-6-phosphate (IGF-II/Man6P) receptors in cultured bovine alveolar macrophages (BAM) was demonstrated by competitive binding studies and crosslinking experiments. Western blotting of protein extracts...
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| Veröffentlicht in: | Molecular and cellular biochemistry 1997-12, Vol.177 (1-2), p.33-45 |
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| creator | Geertz, R Kiess, W Kessler, U Hoeflich, A Tarnok, A Gercken, G |
| description | Expression of insulin-like growth factor-I (IGF-I) receptors and insulin-like growth factor-II/mannose-6-phosphate (IGF-II/Man6P) receptors in cultured bovine alveolar macrophages (BAM) was demonstrated by competitive binding studies and crosslinking experiments. Western blotting of protein extracts from cultured BAM using an anti bovine IGF-II/Man6P receptor antiserum (#66416) confirmed the presence of IGF-II/Man6P receptors on BAM. The effects of IGFs and Man6P on generation of inositol phosphates was measured by HPLC analysis of perchloric acid extracts from myo-[3H]inositol-labelled cultured BAM. IGF-I at nanomolar concentrations and Man6P (10[-8]-10[-3] M) stimulated the accumulation of both Ins(1,4,5)P3 and Ins(1,3,4,5)P4 after 30 sec. IGF-II (up to 2.3 x 10[-8] M) had no significant effect on inositol phosphate accumulation under the same conditions. Both IGFs and Man6P induced a rise in [Ca2+]i in cultured BAM. In addition, using the fluorescent dye SNARF-1/AM we could demonstrate rapid but small IGF-II (10[-9] M) triggered acidification (0.07 pH units) of cultured BAM. Taken together, our results indicate not only the presence of both IGF-I and IGF-II/Man6P receptors on BAM, but also provide evidence of the linkage of the IGF-I receptor to the inositol phosphate system. |
| doi_str_mv | 10.1023/A:1006836631673 |
| format | Article |
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Western blotting of protein extracts from cultured BAM using an anti bovine IGF-II/Man6P receptor antiserum (#66416) confirmed the presence of IGF-II/Man6P receptors on BAM. The effects of IGFs and Man6P on generation of inositol phosphates was measured by HPLC analysis of perchloric acid extracts from myo-[3H]inositol-labelled cultured BAM. IGF-I at nanomolar concentrations and Man6P (10[-8]-10[-3] M) stimulated the accumulation of both Ins(1,4,5)P3 and Ins(1,3,4,5)P4 after 30 sec. IGF-II (up to 2.3 x 10[-8] M) had no significant effect on inositol phosphate accumulation under the same conditions. Both IGFs and Man6P induced a rise in [Ca2+]i in cultured BAM. In addition, using the fluorescent dye SNARF-1/AM we could demonstrate rapid but small IGF-II (10[-9] M) triggered acidification (0.07 pH units) of cultured BAM. Taken together, our results indicate not only the presence of both IGF-I and IGF-II/Man6P receptors on BAM, but also provide evidence of the linkage of the IGF-I receptor to the inositol phosphate system.</description><identifier>ISSN: 0300-8177</identifier><identifier>EISSN: 1573-4919</identifier><identifier>DOI: 10.1023/A:1006836631673</identifier><identifier>PMID: 9450643</identifier><language>eng</language><publisher>Netherlands: Springer Nature B.V</publisher><subject>Acidification ; Animals ; Binding Sites ; Calcium - metabolism ; Cattle ; Cells, Cultured ; Hydrogen-Ion Concentration - drug effects ; Inositol Phosphates - biosynthesis ; Inositol Phosphates - metabolism ; Insulin-Like Growth Factor I - metabolism ; Insulin-Like Growth Factor I - pharmacology ; Insulin-Like Growth Factor II - metabolism ; Insulin-Like Growth Factor II - pharmacology ; Insulin-like growth factors ; Liquid chromatography ; Macrophages, Alveolar - metabolism ; Mannosephosphates - pharmacology ; Proteins ; Receptor, IGF Type 1 - biosynthesis ; Receptor, IGF Type 2 - biosynthesis</subject><ispartof>Molecular and cellular biochemistry, 1997-12, Vol.177 (1-2), p.33-45</ispartof><rights>Kluwer Academic Publishers 1997</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c1263-77d3c04d8158f2dbe29c2de8a678941447d949ceaa20003c74bd161b9ff069073</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,776,780,27903,27904</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/9450643$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Geertz, R</creatorcontrib><creatorcontrib>Kiess, W</creatorcontrib><creatorcontrib>Kessler, U</creatorcontrib><creatorcontrib>Hoeflich, A</creatorcontrib><creatorcontrib>Tarnok, A</creatorcontrib><creatorcontrib>Gercken, G</creatorcontrib><title>Expression of IGF receptors on alveolar macrophages: IGF-induced changes in InsPi formation, [Ca2+]i, and pHi</title><title>Molecular and cellular biochemistry</title><addtitle>Mol Cell Biochem</addtitle><description>Expression of insulin-like growth factor-I (IGF-I) receptors and insulin-like growth factor-II/mannose-6-phosphate (IGF-II/Man6P) receptors in cultured bovine alveolar macrophages (BAM) was demonstrated by competitive binding studies and crosslinking experiments. Western blotting of protein extracts from cultured BAM using an anti bovine IGF-II/Man6P receptor antiserum (#66416) confirmed the presence of IGF-II/Man6P receptors on BAM. The effects of IGFs and Man6P on generation of inositol phosphates was measured by HPLC analysis of perchloric acid extracts from myo-[3H]inositol-labelled cultured BAM. IGF-I at nanomolar concentrations and Man6P (10[-8]-10[-3] M) stimulated the accumulation of both Ins(1,4,5)P3 and Ins(1,3,4,5)P4 after 30 sec. IGF-II (up to 2.3 x 10[-8] M) had no significant effect on inositol phosphate accumulation under the same conditions. Both IGFs and Man6P induced a rise in [Ca2+]i in cultured BAM. In addition, using the fluorescent dye SNARF-1/AM we could demonstrate rapid but small IGF-II (10[-9] M) triggered acidification (0.07 pH units) of cultured BAM. Taken together, our results indicate not only the presence of both IGF-I and IGF-II/Man6P receptors on BAM, but also provide evidence of the linkage of the IGF-I receptor to the inositol phosphate system.</description><subject>Acidification</subject><subject>Animals</subject><subject>Binding Sites</subject><subject>Calcium - metabolism</subject><subject>Cattle</subject><subject>Cells, Cultured</subject><subject>Hydrogen-Ion Concentration - drug effects</subject><subject>Inositol Phosphates - biosynthesis</subject><subject>Inositol Phosphates - metabolism</subject><subject>Insulin-Like Growth Factor I - metabolism</subject><subject>Insulin-Like Growth Factor I - pharmacology</subject><subject>Insulin-Like Growth Factor II - metabolism</subject><subject>Insulin-Like Growth Factor II - pharmacology</subject><subject>Insulin-like growth factors</subject><subject>Liquid chromatography</subject><subject>Macrophages, Alveolar - metabolism</subject><subject>Mannosephosphates - pharmacology</subject><subject>Proteins</subject><subject>Receptor, IGF Type 1 - 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Academic</collection><jtitle>Molecular and cellular biochemistry</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Geertz, R</au><au>Kiess, W</au><au>Kessler, U</au><au>Hoeflich, A</au><au>Tarnok, A</au><au>Gercken, G</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Expression of IGF receptors on alveolar macrophages: IGF-induced changes in InsPi formation, [Ca2+]i, and pHi</atitle><jtitle>Molecular and cellular biochemistry</jtitle><addtitle>Mol Cell Biochem</addtitle><date>1997-12</date><risdate>1997</risdate><volume>177</volume><issue>1-2</issue><spage>33</spage><epage>45</epage><pages>33-45</pages><issn>0300-8177</issn><eissn>1573-4919</eissn><abstract>Expression of insulin-like growth factor-I (IGF-I) receptors and insulin-like growth factor-II/mannose-6-phosphate (IGF-II/Man6P) receptors in cultured bovine alveolar macrophages (BAM) was demonstrated by competitive binding studies and crosslinking experiments. Western blotting of protein extracts from cultured BAM using an anti bovine IGF-II/Man6P receptor antiserum (#66416) confirmed the presence of IGF-II/Man6P receptors on BAM. The effects of IGFs and Man6P on generation of inositol phosphates was measured by HPLC analysis of perchloric acid extracts from myo-[3H]inositol-labelled cultured BAM. IGF-I at nanomolar concentrations and Man6P (10[-8]-10[-3] M) stimulated the accumulation of both Ins(1,4,5)P3 and Ins(1,3,4,5)P4 after 30 sec. IGF-II (up to 2.3 x 10[-8] M) had no significant effect on inositol phosphate accumulation under the same conditions. Both IGFs and Man6P induced a rise in [Ca2+]i in cultured BAM. In addition, using the fluorescent dye SNARF-1/AM we could demonstrate rapid but small IGF-II (10[-9] M) triggered acidification (0.07 pH units) of cultured BAM. Taken together, our results indicate not only the presence of both IGF-I and IGF-II/Man6P receptors on BAM, but also provide evidence of the linkage of the IGF-I receptor to the inositol phosphate system.</abstract><cop>Netherlands</cop><pub>Springer Nature B.V</pub><pmid>9450643</pmid><doi>10.1023/A:1006836631673</doi><tpages>13</tpages></addata></record> |
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| ispartof | Molecular and cellular biochemistry, 1997-12, Vol.177 (1-2), p.33-45 |
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| language | eng |
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| source | MEDLINE; Springer Nature - Complete Springer Journals |
| subjects | Acidification Animals Binding Sites Calcium - metabolism Cattle Cells, Cultured Hydrogen-Ion Concentration - drug effects Inositol Phosphates - biosynthesis Inositol Phosphates - metabolism Insulin-Like Growth Factor I - metabolism Insulin-Like Growth Factor I - pharmacology Insulin-Like Growth Factor II - metabolism Insulin-Like Growth Factor II - pharmacology Insulin-like growth factors Liquid chromatography Macrophages, Alveolar - metabolism Mannosephosphates - pharmacology Proteins Receptor, IGF Type 1 - biosynthesis Receptor, IGF Type 2 - biosynthesis |
| title | Expression of IGF receptors on alveolar macrophages: IGF-induced changes in InsPi formation, [Ca2+]i, and pHi |
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