The elasticity of individual titin PEVK exons measured by single molecule atomic force microscopy.
The I-band region of the giant muscle protein titin contains a large domain enriched for the amino acids proline, glutamate, valine, and lysine and is denoted the PEVK domain. The PEVK domain of titin encodes a random coil shown to be an important factor in the passive elasticity of titin. Muscle-specific splicing of 116 PEVK exons encodes this domain. It has been proposed that proline contents determine the elasticity of the PEVK polypeptide, where the individual exons code for "flexibility cassettes." To test this hypothesis, we have measured the elasticity of three distinct polypeptides encoded by individual PEVK exons (161, 120 and 184) that varied greatly in their proline contents (7, 14, and 37% respectively) and total PEVK contents (55, 70, and 87%). We used single molecule atomic force microscopy techniques to measure the persistence length, p, of the engineered PEVK proteins. Surprisingly, all three exons 161, 120, and 184 coded for proteins with similar values of persistence length, p = 0.92 +/- 0.38, 0.89 +/- 0.42, and 0.98 +/- 0.4 nm, respectively. We conclude that the PEVK exons encode polypeptides of similar elastic properties, unrelated to their total PEVK contents. Hence, alternative splicing solely adjusts the length of the PEVK domain of titin.
Sarkar
Atom
A
Department of Biological Sciences, Columbia University, New York, New York 10027, USA.
Caamano
Sofia
S
Fernandez
Julio M
JM
D017398:Alternative Splicing; D000595:Amino Acid Sequence; D064211:Connectin; D004548:Elasticity; D005091:Exons; D006801:Humans; D018625:Microscopy, Atomic Force; D009124:Muscle Proteins / Q000737:chemistry* / Q000235:genetics / Q000502:physiology; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry; D015202:Protein Engineering; D011494:Protein Kinases / Q000737:chemistry* / Q000235:genetics / Q000502:physiology; D012091:Repetitive Sequences, Nucleic Acid; D013329:Structure-Activity Relationship
D064211:Connectin; D009124:Muscle Proteins; D010455:Peptides; C577285:TTN protein, human; D011494:Protein Kinases