Study of human laryngeal muscle protein using two-dimensional electrophoresis and mass spectrometry

• Published in: Proteomics (Weinheim) 2003-07, Vol.3 (7), p.1325-1334
• Main Authors: Li, Zhao-Bo, Lehar, Mohamed, Braga, Natasha, Westra, William, Liu, Li-Hong, Flint, Paul W.
• Format: Article
• Language: English
• Subjects: Databases as Topic; Electrophoresis, Gel, Two-Dimensional - methods; Electrophoresis, Polyacrylamide Gel; Human laryngeal muscle; Humans; Hydrogen-Ion Concentration; Isoelectric Focusing - methods; Laryngeal Muscles - metabolism; Laryngeal Neoplasms; Protein Biosynthesis; Proteome; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization - methods; Time Factors; Two-dimensional gel electrophoresis
• ISSN: 1615-9853; 1615-9861
• DOI: 10.1002/pmic.200300454

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.