Troponin-T is a calcium-binding protein in insect muscle: in vivo phosphorylation, muscle-specific isoforms and developmental profile in Drosophila melanogaster.
Two sets of muscle polypeptides showing calcium-binding capacity and intense labelling in vivo with 32P were purified and characterized from Drosophila melanogaster adult extracts. The polypeptides exhibit crossed immunoreactivity and share similar biochemical properties such as those involved in purification. They have been identified as isoforms of troponin-T (TnT) by sequence analysis of a cDNA clone isolated from an embryonic library. The two sets of TnT polypeptides correspond to the fibrillar and non-fibrillar muscle isoforms, respectively. The non-fibrillar muscle isoforms separate into two bands which are differentially expressed during development. Analysis of TnT isoforms in bee thoraces indicates that the expression of the fibrillar muscle isoform correlates with the acquisition of functional flight capability. In vivo labelling experiments reveal that the two TnT sets are readily phosphorylated. The Drosophila TnTs show calcium-binding properties by three different types of assays. Our results suggest that this property could be specific to insect TnTs and may be related to the long, extremely acidic polyglutamic carboxy-terminus present in these polypeptides, which does not occur in non-arthropod TnTs.
Domingo
A
A
Departamento de Bioquímica de la UAM, Spain.
González-Jurado
J
J
Maroto
M
M
Díaz
C
C
Vinós
J
J
Carrasco
C
C
Cervera
M
M
Marco
R
R
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D000911:Antibodies, Monoclonal; D002135:Calcium-Binding Proteins / Q000378:metabolism*; D003429:Cross Reactions; D004331:Drosophila melanogaster / Q000254:growth & development / Q000378:metabolism*; D006110:Grasshoppers; D015151:Immunoblotting; D019476:Insect Proteins / Q000378:metabolism*; D007536:Isomerism; D024510:Muscle Development; D009132:Muscles / Q000737:chemistry / Q000378:metabolism; D010766:Phosphorylation; D014336:Troponin / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D020107:Troponin T
D000596:Amino Acids; D000911:Antibodies, Monoclonal; D002135:Calcium-Binding Proteins; D019476:Insect Proteins; D014336:Troponin; D020107:Troponin T