Smith R K, Zunino L, Webbon P M, Heinegård D
Department of Farm Animal and Equine Medicine and Surgery, Royal Veterinary College, Hatfield, Hertfordshire, UK.
Matrix Biol. 1997 Nov;16(5):255-71. doi: 10.1016/s0945-053x(97)90014-7.
A protein prominent in guanidine hydrochloride extracts of adult bovine and equine digital flexor tendons was confirmed to be Cartilage Oligomeric Matrix Protein (COMP) by non-reducing and reducing SDS-PAGE, reaction with rabbit anti-COMP polyclonal antiserum on Western blots, trypsin digestion followed by HPLC on a C2/C18 column, and identification of COMP mRNA from tendon on Northern blots. Immunohistochemistry and Western blots of extracts showed COMP to be present in all regions of digital flexor tendons. Equine tendon COMP was purified by ion exchange chromatography and gel filtration and used in a heterologous inhibition ELISA to quantify COMP in equine digital flexor tendons at different ages, and in other tendons and ligaments. Mean COMP levels in digital flexor tendon were approximately 2-5mg/g wet weight, but they showed a large variation. Levels were low in neonatal tendon but rose rapidly during growth, with the metacarpal (tensional) superficial digital flexor tendon having the highest levels (approximately 10mg/g wet weight). Levels subsequently declined in this region, while in areas which experience a variable amount of compression, levels increased less but then remained constant. Extensor tendons and collateral ligaments, which experience less loading in vivo, had levels similar to those in neonatal tendon. COMP was identified in scarred skin and granulation tissue but not in normal skin, chronic fibrosis, or a fibrosarcomatous skin growth. A unilateral non-weight-bearing growing animal contained three to six times more COMP in the weight-bearing digital flexor tendons compared to the paralyzed limb, while the extensor tendons had similar amounts in both limbs. With the recent discovery of a COMP gene mutation causing pseudoachondroplasia (Hecht et al., 1995), in which lax tendons and ligaments are a feature, the present data suggest that COMP is synthesized in response to, and is necessary for tendon to resist, load.
通过非还原和还原SDS - PAGE、在蛋白质免疫印迹上与兔抗软骨寡聚基质蛋白(COMP)多克隆抗血清反应、胰蛋白酶消化后在C2/C18柱上进行高效液相色谱分析以及在Northern印迹上从肌腱中鉴定COMP mRNA,证实了成年牛和马的指屈肌腱盐酸胍提取物中的一种主要蛋白质为软骨寡聚基质蛋白(COMP)。提取物的免疫组织化学和蛋白质免疫印迹显示COMP存在于指屈肌腱的所有区域。马肌腱COMP通过离子交换色谱和凝胶过滤进行纯化,并用于异源抑制酶联免疫吸附测定,以定量不同年龄马指屈肌腱、其他肌腱和韧带中的COMP。指屈肌腱中COMP的平均水平约为2 - 5mg/g湿重,但存在较大差异。新生肌腱中的水平较低,但在生长过程中迅速升高,掌骨(受张力)浅指屈肌腱中的水平最高(约10mg/g湿重)。该区域的水平随后下降,而在经历不同程度压缩的区域,水平升高较少但随后保持稳定。在体内承受负荷较少的伸肌腱和侧副韧带,其水平与新生肌腱相似。在瘢痕皮肤和肉芽组织中鉴定出COMP,但在正常皮肤、慢性纤维化或纤维肉瘤性皮肤生长中未鉴定出。一只单侧非负重生长的动物,其负重的指屈肌腱中的COMP含量比瘫痪肢体中的多三到六倍,而伸肌腱在两个肢体中的含量相似。随着最近发现一种导致假性软骨发育不全的COMP基因突变(赫克特等人,1995年),其中肌腱和韧带松弛是一个特征,目前的数据表明COMP是在对负荷作出反应时合成的,并且是肌腱抵抗负荷所必需的。