Study of human laryngeal muscle protein using two-dimensional electrophoresis and mass spectrometry.
Proteomic analysis was performed to construct a protein database for human laryngeal muscle. Thyroarytenoid (TA) muscle specimens were obtained from six post mortem cases within 24 h of death. Isoelectric focusing was performed by using immobilized pH gradient strips followed by 12% sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Silver stained gels were then analyzed using PDQuest software to locate, quantify and match spots. Proteins were identified by matrix-assisted laser desorption/ionization-mass spectrometry on the basis of peptide mass fingerprinting following in-gel digestion with trypsin. Comparison of protein distribution between broad and narrow pH range gels demonstrated that 75% of all protein spots from human TA muscle were located within the pH range 5-8, and between mass 15-120 kDa. Based on peptide mass fingerprinting, 75 proteins were identified and classified into six functional groups. These include membrane proteins (8.5%), cytoskeletal and myofibrillar proteins (14.6%), energy production proteins (28%), proteins associated with stress responses (8.5%), and protein associated with transcription regulation (10.9%). Approximately one-third (29%) were categorized as "other proteins". This data provides an initial reference map for comparative studies of protein expression in human and laryngeal muscle. Further development of this database will provide a valuable resource for molecular analysis of normal and pathologic conditions affecting human striated muscle.
Li
Zhao-Bo
ZB
Department of Otolaryngology-Head and Neck Surgery, Johns Hopkins University School of Medicine, 601 N. Caroline Street, Baltimore, MD 21287, USA.
Lehar
Mohamed
M
Braga
Natasha
N
Westra
William
W
Liu
Li-Hong
LH
Flint
Paul W
PW
D019992:Databases as Topic; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007525:Isoelectric Focusing / Q000379:methods*; D007821:Laryngeal Muscles / Q000378:metabolism*; D007822:Laryngeal Neoplasms; D014176:Protein Biosynthesis; D020543:Proteome; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D013997:Time Factors
D020543:Proteome