Patel Sunita R, Saide Judith D
Department of Physiology and Biophysics, Boston University School of Medicine, Boston, MA 02118, USA.
J Muscle Res Cell Motil. 2005;26(4-5):213-24. doi: 10.1007/s10974-005-9012-y. Epub 2005 Nov 4.
Stretchin-klp is a newly described protein in Drosophila indirect flight muscles (IFM) that migrates on SDS gels as two distinct components of approximately 225 and 231 kD. Although the larger isoform is IFM specific, the smaller stretchin-klp isoform is expressed not only in IFM, but also in wild-type tissues of the adult head, abdomen and thorax from which the IFM has been removed. It is not detected, however, in jump or leg muscles. Probes derived from a cDNA encoding part of stretchin-klp hybridize with a 6.7 kb mRNA. Stretchin-klp is one of several putative products of the Stretchin-Myosin light chain kinase gene and is predicted to have multiple immunoglobulin domains arranged in tandem pairs separated by variable length spacers. Polyclonal antibodies directed against the expressed peptide of the stretchin-klp cDNA label the IFM myofibril A-band, though not its central and lateral regions. Analyses of IFM mutants indicate that the larger stretchin-klp isoform is myosin dependent. Although the normal adult myosin filament or the 'headless' myosin rod is sufficient for accumulation of both the large and small stretchin-klp isoforms, loss of myosin, or substitution of the adult rod with an embryonic one in IFM prevents the larger isoform from being formed or stabilized. During development stretchin-klp is first detected at pupal stage p8, when myofibrils are being constructed. These studies suggest that this newly identified protein is a major component of the Drosophila IFM thick filament.
伸展蛋白 - 肌球蛋白轻链激酶(Stretchin-klp)是果蝇间接飞行肌(IFM)中一种新发现的蛋白质,它在SDS凝胶上迁移时呈现为两条不同的条带,分子量约为225 kD和231 kD。虽然较大的同工型是IFM特有的,但较小的伸展蛋白 - klp同工型不仅在IFM中表达,还在去除了IFM的成年果蝇头部、腹部和胸部的野生型组织中表达。然而,在跳跃肌或腿部肌肉中未检测到它。从编码伸展蛋白 - klp部分序列的cDNA衍生的探针与一个6.7 kb的mRNA杂交。伸展蛋白 - klp是伸展蛋白 - 肌球蛋白轻链激酶基因的几种推定产物之一,预计具有多个免疫球蛋白结构域,这些结构域以可变长度的间隔区串联成对排列。针对伸展蛋白 - klp cDNA表达肽的多克隆抗体标记IFM肌原纤维的A带,但不标记其中心和外侧区域。对IFM突变体的分析表明,较大的伸展蛋白 - klp同工型依赖于肌球蛋白。虽然正常的成年肌球蛋白丝或“无头”肌球蛋白杆足以使大小两种伸展蛋白 - klp同工型积累,但肌球蛋白的缺失,或在IFM中用胚胎肌球蛋白杆替代成年肌球蛋白杆,会阻止较大同工型的形成或稳定。在发育过程中,伸展蛋白 - klp最早在蛹期p8被检测到,此时肌原纤维正在构建。这些研究表明,这种新鉴定的蛋白质是果蝇IFM粗丝的主要成分。