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转谷氨酰胺酶,即因子ⅫIA,存在于关节软骨细胞中。

The transglutaminase, Factor XIIIA, is present in articular chondrocytes.

作者信息

Rosenthal A K, Masuda I, Gohr C M, Derfus B A, Le M

机构信息

Zablocki VAMC, Milwaukee, WI 53295-1000, USA.

出版信息

Osteoarthritis Cartilage. 2001 Aug;9(6):578-81. doi: 10.1053/joca.2000.0423.

Abstract

OBJECTIVES

The transglutaminase (TGase) family includes seven different enzymes that catalyse a protein cross-linking reaction resulting in structural and functional alterations in substrate proteins. TGase activity is easily measureable in mature articular cartilage where it may contribute to CPPD deposition disease through its actions on growth factors, crystal components or extracellular matrix proteins. In contrast, low levels of TGase activity are found in chondrocytes from young animals. We previously demonstrated type II TGase protein in articular chondrocytes. Earlier work also suggested the presence of another form of TGase in chondrocytes. We sought to determine if articular chondrocytes contain the TGase, Factor XIIIA (FXIIIA).

METHODS

Western blots with FXIIIA antibody were used to detect FXIIIA in young and old porcine articular chondrocytes and articular cartilage vesicles (ACVs). The presence of FXIIIA mRNA was confirmed by RT-PCR.

RESULTS

Old chondrocyte conditioned medium, cytosol, and membrane fractions contained FXIIIA protein on Western blots, while less FXIIIA was detectable in cell fractions or media from young chondrocytes. ACVs also contained FXIIIA. FXIIIA mRNA was demonstrated by PCR in old and young chondrocytes.

CONCLUSIONS

FXIIIA is present in articular chondrocytes. FXIIIA levels correlate with TGase activity in chondrocytes. The presence of two forms of TGase in articular chondrocytes suggest an important function for this enzyme family in articular cartilage.

摘要

目的

转谷氨酰胺酶(TGase)家族包含七种不同的酶,它们催化蛋白质交联反应,导致底物蛋白的结构和功能改变。TGase活性在成熟关节软骨中易于测量,它可能通过对生长因子、晶体成分或细胞外基质蛋白的作用,导致焦磷酸钙沉积病。相比之下,在幼龄动物的软骨细胞中发现TGase活性水平较低。我们之前在关节软骨细胞中证实了II型TGase蛋白的存在。早期研究还表明软骨细胞中存在另一种形式的TGase。我们试图确定关节软骨细胞中是否含有转谷氨酰胺酶因子XIIIA(FXIIIA)。

方法

使用FXIIIA抗体进行蛋白质免疫印迹,以检测幼年和老年猪关节软骨细胞及关节软骨小泡(ACV)中的FXIIIA。通过逆转录聚合酶链反应(RT-PCR)确认FXIIIA mRNA的存在。

结果

蛋白质免疫印迹显示,老年软骨细胞条件培养基、细胞溶质和膜组分中含有FXIIIA蛋白,而幼年软骨细胞的细胞组分或培养基中可检测到的FXIIIA较少。ACV中也含有FXIIIA。PCR证实幼年和老年软骨细胞中均存在FXIIIA mRNA。

结论

关节软骨细胞中存在FXIIIA。FXIIIA水平与软骨细胞中的TGase活性相关。关节软骨细胞中两种形式TGase的存在表明该酶家族在关节软骨中具有重要功能。

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