BIOCHEMICAL, STRUCTURAL, AND TRANSGLUTAMINASE SUBSTRATE PROPERTIES OF HUMAN LORICRIN, THE MAJOR EPIDERMAL CORNIFIED CELL-ENVELOPE PROTEIN
BIOCHEMICAL, STRUCTURAL, AND TRANSGLUTAMINASE SUBSTRATE PROPERTIES OF HUMAN LORICRIN, THE MAJOR EPIDERMAL CORNIFIED CELL-ENVELOPE PROTEIN
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DOI：
10.1074/jbc.270.44.26382
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发表时间：
1995-11-03
影响因子：
4.8
通讯作者：
STEINERT, PM
中科院分区：
文献类型：
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作者：
CANDI, E;MELINO, G;STEINERT, PM
Loricrin是终末分化的表皮角质形成细胞角化细胞膜的主要蛋白质，起着物理屏障的作用，为了了解其在角化细胞囊膜中的性质和作用，我们从细菌中表达了人Loricrin，并对其进行了纯化。我们还从新生小鼠的表皮中分离出了氯氯丙烷。圆二色谱和荧光光谱分析表明，在体内的小鼠和细菌表达的人三氯鸟蛋白不具有α或β结构，但在溶液中具有与其多种酪氨酸相关的有组织结构，并且可以被盐酸胍或温度可逆变性。表皮分化过程中表达的谷氨酰胺转氨酶(TGase)1、2和3酶在体外以氯丙烷为完全底物，但交联物的类型不同。TGase 3反应主要通过形成链内交联物而有利于某些赖氨酸和谷氨酸，而TGase 1通过涉及不同赖氨酸和谷氨酸的链间交联物形成大部分大的寡聚复合体。体外使用的谷氨酸和赖氨酸加在一起，与体内使用的几乎完全相同。这些数据支持一种假说，即TGase 1和TGase 3在体内氯氯菊酯的交联过程中都发挥着基本的和互补的作用。TGase 1不能使氯化蛋白交联，这可能解释了板层鱼鳞病的表型，这是一种由TGase 1基因突变引起的疾病。
Loricrin is the major protein of the cornified cell envelope of terminally differentiated epidermal keratinocytes which functions as a physical barrier, In order to understand its properties and role in cornified cell envelope, we have expressed human loricrin from a full-length cDNA clone in bacteria and purified it to homogeneity. We have also isolated loricrin from newborn mouse epidermis. By circular dichroism and fluorescence spectroscopy, the in vivo mouse and bacterially expressed human loricrins possess no alpha or beta structure but have some organized structure in solution associated with their multiple tyrosines and can be reversibly denatured by either guanidine hydrochloride or temperature. The transglutaminase (TGase) 1, 2, and 3 enzymes expressed during epidermal differentiation utilized loricrin in vitro as a complete substrate, but the types of cross-linking were different. The TGase 3 reaction favored certain lysines and glutamines by forming mostly intrachain cross-links, whereas TGase 1 formed mostly large oligomeric complexes by interchain crosslinks involving different lysines and glutamines. Together, the glutamines and lysines used in vitro are almost identical to those seen in vivo. The data support a hypothesis for the essential and complementary roles of both TGase 1 and TGase 3 in cross-linking of loricrin in vivo. Failure to cross-link loricrin by TGase 1 may explain the phenotype of lamellar ichthyosis, a disease caused by mutations in the TGase 1 gene.