Hierarchical extensibility in the PEVK domain of skeletal-muscle titin.
Titin is the main determinant of passive muscle force. Physiological extension of titin derives largely from its PEVK (Pro-Glu-Val-Lys) domain, which has a different length in different muscle types. Here we characterized the elasticity of the full-length, human soleus PEVK domain by mechanically manipulating its contiguous, recombinant subdomain segments: an N-terminal (PEVKI), a middle (PEVKII), and a C-terminal (PEVKIII) one third. Measurement of the apparent persistence lengths revealed a hierarchical arrangement according to local flexibility: the N-terminal PEVKI is the most rigid and the C-terminal PEVKIII is the most flexible segment within the domain. Immunoelectron microscopy supported the hierarchical extensibility within the PEVK domain. The effective persistence lengths decreased as a function of ionic strength, as predicted by the Odijk-Skolnick-Fixman model of polyelectrolyte chains. The ionic strength dependence of persistence length was similar in all segments, indicating that the residual differences in the elasticity of the segments derive from nonelectrostatic mechanisms.
Nagy
A
A
Department of Biophysics, University of Pécs, Faculty of Medicine, Pécs, H-7624 Hungary.
Grama
L
L
Huber
T
T
Bianco
P
P
Trombitás
K
K
Granzier
H L
HL
Kellermayer
M S Z
MS
D020816:Amino Acid Motifs; D001703:Biophysics / Q000379:methods; D003001:Cloning, Molecular; D064211:Connectin; D018076:DNA, Complementary / Q000378:metabolism; D015723:Gene Library; D006801:Humans; D007477:Ions; D016253:Microscopy, Immunoelectron; D009119:Muscle Contraction; D009124:Muscle Proteins / Q000737:chemistry*; D018482:Muscle, Skeletal / Q000378:metabolism*; D011494:Protein Kinases / Q000737:chemistry*; D017434:Protein Structure, Tertiary; D012518:Sarcomeres / Q000378:metabolism; D013053:Spectrophotometry; D055672:Static Electricity; D013314:Stress, Mechanical
D064211:Connectin; D018076:DNA, Complementary; D007477:Ions; D009124:Muscle Proteins; C577285:TTN protein, human; D011494:Protein Kinases