Identification and characterization of homologues of vertebrate beta-thymosin in the marine mollusk Aplysia californica.
The beta-thymosins have been known as actin-sequestering proteins, but now are recognized as molecules with multiple and diverse intracellular and extracellular functions. Two closely related proteins, beta-thymosin(His) and beta-thymosin(Gln), have been de novo sequenced by top-down mass spectrometry in the common neurobiology model, Aplysia californica. As determined by nanoelectrospray quadrupole-enhanced Fourier-Transform mass spectrometry with collisionally activated and electron-capture dissociations, both of these Aplysia beta-thymosins are acetylated and differ by a single residue in the central actin-binding domain. Profiling of individual cells and tissue by matrix-assisted laser desorption/ionization mass spectrometry reveals that these proteins are widely expressed in the Aplysia central nervous system, including in individual identified neurons, neuronal clusters, nerves and connective tissues. Newly identified beta-thymosin(His) and beta-thymosin(Gln) are also detected by mass spectrometry in hemolymph, and in releasates collected from whole ganglia. When applied exogenously, beta-thymosin proteins, purified from nerve cell extract, support the anchoring of neurons, and increase neurite sprouting and total neurite outgrowth in culture. These positive effects on neurite regeneration in cell culture suggest that the beta-thymosin proteins have an extracellular function in the central nervous system of Aplysia californica.
Romanova
Elena V
EV
Department of Chemistry, University of Illinois and Beckman Institute for Advanced Science and Technology, Urbana, IL 61801, USA.
Roth
Michael J
MJ
Rubakhin
Stanislav S
SS
Jakubowski
Jennifer A
JA
Kelley
Wayne P
WP
Kirk
Mark D
MD
Kelleher
Neil L
NL
Sweedler
Jonathan V
JV
D000595:Amino Acid Sequence; D000818:Animals; D001048:Aplysia / Q000737:chemistry* / Q000235:genetics; D001681:Biological Assay; D002478:Cells, Cultured; D002851:Chromatography, High Pressure Liquid; D004594:Electrophysiology; D045604:Extracellular Fluid / Q000737:chemistry; D008839:Microelectrodes; D008969:Molecular Sequence Data; D036103:Nanotechnology; D016501:Neurites / Q000502:physiology; D009473:Neuronal Plasticity / Q000502:physiology; D021241:Spectrometry, Mass, Electrospray Ionization; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013947:Thymosin / Q000032:analysis / Q000737:chemistry* / Q000235:genetics; D014020:Tissue Extracts / Q000737:chemistry
D014020:Tissue Extracts; D013947:Thymosin