{"title":"[3H]转谷氨酰胺酶抑制剂对皮肤伤口愈合的影响。","authors":"J M Bowness","doi":"10.1139/o82-096","DOIUrl":null,"url":null,"abstract":"<p><p>Punch wounds (3 mm) were made in the skin of rats and the animals were killed after 1 or 3 days. Plugs (4 mm) of wounded and unwounded skin were incubated in vitro with [3H]fucose. The labelled plugs were homogenized and subjected to sequential extraction with buffered salt solutions, ethanol-ether, and 8 M urea - 50 mM dithiothreitol (DTT). Nondialysable counts in the extracts and insoluble residue were determined and the incorporation of label by wounded and unwounded skin plugs was compared. Wound plugs showed a greater total incorporation of [3H]fucose. In addition, a greater proportion of [3H]fucose was found in the urea-DTT extracts. The highest specific activity (disintegrations per minute [3H]fucose per milligram dry weight) was found in a finely dispersed precipitate, sedimenting at 10000 x g but not at 1000 x g. The transglutaminase inhibitors aminoacetonitrile and dansyl cadaverine were found to increase the extractability of a portion of the material which incorporated [3H]fucose without affecting the total incorporation. These results show that healing wounds have an increased biosynthetic capacity for an insoluble fucosylated glycoprotein fraction and they suggest that transglutaminase is necessary to make this fraction fully insoluble.</p>","PeriodicalId":9508,"journal":{"name":"Canadian journal of biochemistry","volume":"60 8","pages":"777-81"},"PeriodicalIF":0.0000,"publicationDate":"1982-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1139/o82-096","citationCount":"5","resultStr":"{\"title\":\"[3H]Fucose incorporation by healing skin wounds and the effect of transglutaminase inhibitors.\",\"authors\":\"J M Bowness\",\"doi\":\"10.1139/o82-096\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>Punch wounds (3 mm) were made in the skin of rats and the animals were killed after 1 or 3 days. Plugs (4 mm) of wounded and unwounded skin were incubated in vitro with [3H]fucose. The labelled plugs were homogenized and subjected to sequential extraction with buffered salt solutions, ethanol-ether, and 8 M urea - 50 mM dithiothreitol (DTT). Nondialysable counts in the extracts and insoluble residue were determined and the incorporation of label by wounded and unwounded skin plugs was compared. Wound plugs showed a greater total incorporation of [3H]fucose. In addition, a greater proportion of [3H]fucose was found in the urea-DTT extracts. The highest specific activity (disintegrations per minute [3H]fucose per milligram dry weight) was found in a finely dispersed precipitate, sedimenting at 10000 x g but not at 1000 x g. The transglutaminase inhibitors aminoacetonitrile and dansyl cadaverine were found to increase the extractability of a portion of the material which incorporated [3H]fucose without affecting the total incorporation. These results show that healing wounds have an increased biosynthetic capacity for an insoluble fucosylated glycoprotein fraction and they suggest that transglutaminase is necessary to make this fraction fully insoluble.</p>\",\"PeriodicalId\":9508,\"journal\":{\"name\":\"Canadian journal of biochemistry\",\"volume\":\"60 8\",\"pages\":\"777-81\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"1982-08-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://sci-hub-pdf.com/10.1139/o82-096\",\"citationCount\":\"5\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Canadian journal of biochemistry\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://doi.org/10.1139/o82-096\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Canadian journal of biochemistry","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1139/o82-096","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 5
摘要
在大鼠皮肤上做3mm的穿孔,1 ~ 3天后处死。损伤和未损伤皮肤(4mm)用[3H]聚焦体外培养。将标记好的桥塞均质,用缓冲盐溶液、乙醇醚和8 M尿素- 50 mM二硫代苏糖醇(DTT)进行顺序提取。测定了提取物中不可透析计数和不溶性残留物计数,并比较了损伤和未损伤皮塞纳入标签的情况。伤口塞显示更多的[3H]病灶。此外,尿素- dtt提取物中发现了更大比例的[3H]焦点。最高的比活性(每毫克干重每分钟[3H]焦的崩解量)是在精细分散的沉淀物中发现的,沉淀在10000 x g,而不是1000 x g。转谷氨酰胺酶抑制剂氨基乙腈和丹酰尸胺被发现增加了含有[3H]焦的部分物质的可提取率,而不影响总掺入量。这些结果表明,愈合的伤口具有增加的不溶性集中糖蛋白部分的生物合成能力,并且表明转谷氨酰胺酶是使该部分完全不溶的必要条件。
[3H]Fucose incorporation by healing skin wounds and the effect of transglutaminase inhibitors.
Punch wounds (3 mm) were made in the skin of rats and the animals were killed after 1 or 3 days. Plugs (4 mm) of wounded and unwounded skin were incubated in vitro with [3H]fucose. The labelled plugs were homogenized and subjected to sequential extraction with buffered salt solutions, ethanol-ether, and 8 M urea - 50 mM dithiothreitol (DTT). Nondialysable counts in the extracts and insoluble residue were determined and the incorporation of label by wounded and unwounded skin plugs was compared. Wound plugs showed a greater total incorporation of [3H]fucose. In addition, a greater proportion of [3H]fucose was found in the urea-DTT extracts. The highest specific activity (disintegrations per minute [3H]fucose per milligram dry weight) was found in a finely dispersed precipitate, sedimenting at 10000 x g but not at 1000 x g. The transglutaminase inhibitors aminoacetonitrile and dansyl cadaverine were found to increase the extractability of a portion of the material which incorporated [3H]fucose without affecting the total incorporation. These results show that healing wounds have an increased biosynthetic capacity for an insoluble fucosylated glycoprotein fraction and they suggest that transglutaminase is necessary to make this fraction fully insoluble.