Pandit Sujata G, Govindraj Prasanthi, Sasse Joachim, Neame Peter J, Hassell John R
The Center for Research in Skeletal Development and Pediatric Orthopedics, Shriners Hospital for Children, 12502 North Pine Drive, Tampa, FL 33612, USA.
Biochem J. 2002 Jan 15;361(Pt 2):231-41. doi: 10.1042/0264-6021:3610231.
Point mutations in the human fibroblast growth factor (FGF) receptor 3 gene (Fgfr3) produce a constitutively active receptor, which disrupts chondrocyte differentiation in the growth plate and results in skeletal dysplasias with severe shortening of the limbs. Alternative splicing of the Fgfr3 transcript gives rise to two isoforms, IIIc and IIIb, which vary in their specificity for FGF ligands. We examined the expression of these FGFR3 isoforms in the bovine fetal rib growth plate to determine whether levels of FGFR3 expression are zone-related. Transcripts for both Fgfr3 isoforms are expressed in rib growth plate, with maximum expression in the hypertrophic region and the least expression in the reserve zone. Fgfr3 IIIc is the predominant isoform in the growth plate. Western-blot analysis revealed the presence of full-length FGFR3 (135 kDa) for both isoforms in the reserve zone, a major 98 kDa fragment in all zones and smaller fragments primarily in the hypertrophic zone. Immunostaining localized FGFR3 to the pericellular region of reserve chondrocytes and to the extracellular matrix in the hypertrophic zone. These results suggest that the transmembrane form of FGFR3 increasingly undergoes proteolytic cleavage towards the hypertrophic zone to produce an extracellular-domain fragment of FGFR3, which is present in large amounts in the matrix of hypertrophic cells. These findings suggest a proteolytic regulatory mechanism for FGFR3, whereby Fgfr3 fragments could control availability of FGF for the intact receptor, and by which proteolysis could inactivate the receptor.
人类成纤维细胞生长因子(FGF)受体3基因(Fgfr3)中的点突变产生一种组成型活性受体,其破坏生长板中的软骨细胞分化并导致肢体严重缩短的骨骼发育异常。Fgfr3转录本的可变剪接产生两种异构体,IIIc和IIIb,它们对FGF配体的特异性不同。我们检测了这些FGFR3异构体在牛胎儿肋骨生长板中的表达,以确定FGFR3表达水平是否与区域相关。两种Fgfr3异构体的转录本均在肋骨生长板中表达,在肥大区表达最高,在储备区表达最低。Fgfr3 IIIc是生长板中的主要异构体。蛋白质印迹分析显示,储备区两种异构体均存在全长FGFR3(135 kDa),所有区域均存在一个主要的98 kDa片段,较小的片段主要存在于肥大区。免疫染色将FGFR3定位到储备软骨细胞的细胞周围区域和肥大区的细胞外基质中。这些结果表明,FGFR3的跨膜形式在向肥大区的过程中越来越多地经历蛋白水解切割,以产生FGFR3的细胞外结构域片段,该片段大量存在于肥大细胞的基质中。这些发现提示了一种FGFR3的蛋白水解调节机制,通过该机制,Fgfr3片段可以控制完整受体的FGF可用性,并且蛋白水解可以使受体失活。