Mass spectrometry analyses of rat 2b myosin heavy chain isoform.
We have separated 2b myosin heavy chain (MyHC) isoform from the rat extensor digitorum longus muscle by SDS-PAGE and analyzed it by two subsequent mass spectrometry techniques. After tryptic digestion, the obtained peptides were identified by Matrix-Assisted Laser Desorption/Ionisation reflectron Time of Flight mass spectrometry (MALDI-TOF MS) and sequenced by liquid chromatography tandem mass spectrometry (ESI LC/MS/MS). The analyzed peptides proportionally covered 30 % of the 2b MyHC isoform sequence. The results suggest that the primary structure is identical with the highest probability to a NCBI database record ref|NP_062198.1|, representing the last updated record of rat 2b isoform. Nonetheless, four peptides carrying amino acid substitution(s) in comparison with the NCBI database record were identified.
Zurmanová
J
J
Institute of Physiology AS CR, Prague, Czech Republic.
Maláčová
D
D
Půta
F
F
Novák
P
P
Rícný
J
J
Soukup
T
T
D000595:Amino Acid Sequence; D000818:Animals; D002853:Chromatography, Liquid; D030562:Databases, Protein; D004591:Electrophoresis, Polyacrylamide Gel; D008969:Molecular Sequence Data; D018482:Muscle, Skeletal / Q000737:chemistry*; D018995:Myosin Heavy Chains / Q000737:chemistry* / Q000302:isolation & purification; D024501:Nonmuscle Myosin Type IIB / Q000737:chemistry* / Q000302:isolation & purification; D010449:Peptide Mapping; D011487:Protein Conformation; D051381:Rats; D021241:Spectrometry, Mass, Electrospray Ionization; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D053719:Tandem Mass Spectrometry
D024501:Nonmuscle Myosin Type IIB; C103815:nonmuscle myosin type IIB heavy chain; D018995:Myosin Heavy Chains