Bowness J M, Tarr A H, Wong T
Department of Biochemistry, Faculty of Medicine, University of Manitoba, Winnipeg, Canada.
Biochim Biophys Acta. 1988 Nov 17;967(2):234-40. doi: 10.1016/0304-4165(88)90014-1.
Outer, middle and inner layers from wounded or unwounded rat dorsal skin were separated and extracted first with buffer and then with Triton X-100 and dithiothreitol. The extracts and residues were assayed for transglutaminase activity and tissue transglutaminase antigen. Transglutaminase activities in all skin layers are increased in the period 1-5 days after wounding. Most of the increased activity is in the buffer-soluble fraction in the inner skin layer though there is no corresponding increase in antigen in this fraction. This suggests that there is production of activated soluble tissue transglutaminase in the wounded inner layer. In the 3-5 day wounded outer layer the largest fraction of both activity and antigen is associated with the insoluble residue remaining after extraction with Triton X-100. On DEAE-cellulose chromatography Triton X-100 extracts of the inner layer of wounded skin showed a single major peak of activity, corresponding approximately with rabbit liver transglutaminase; the outer layer showed the same peak plus a different one, eluting at lower salt concentration, which is thought to be epidermal transglutaminase.
分离受伤或未受伤大鼠背部皮肤的外层、中层和内层，先用缓冲液提取，然后用 Triton X - 100 和二硫苏糖醇提取。对提取物和残留物进行转谷氨酰胺酶活性和组织转谷氨酰胺酶抗原检测。受伤后1 - 5天内，所有皮肤层的转谷氨酰胺酶活性均升高。尽管内层皮肤缓冲液可溶部分的抗原没有相应增加，但大部分增加的活性存在于该部分。这表明受伤的内层产生了活化的可溶性组织转谷氨酰胺酶。在受伤3 - 5天的外层，活性和抗原的最大部分与用 Triton X - 100 提取后残留的不溶性部分相关。在 DEAE - 纤维素柱层析上，受伤皮肤内层的 Triton X - 100 提取物显示出一个单一的主要活性峰，大致与兔肝转谷氨酰胺酶相对应；外层显示出相同的峰以及另一个在较低盐浓度下洗脱的不同峰，该峰被认为是表皮转谷氨酰胺酶。
