Document Detail


MALDI tissue imaging of ocular lens alpha-crystallin.
MedLine Citation:
PMID:  16799044     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
PURPOSE: To apply MALDI (matrix-assisted laser desorption ionization) tissue imaging methods to obtaining a profile of the distribution of the lens alpha-crystallins and their modified forms in calf and mature bovine lenses. METHODS: Frozen bovine lenses were cut equatorially at -12 degrees C to -20 degrees C into 10- to 40-microm sections depending on lens age. Tissue sections were mounted onto MALDI sample plates by ethanol soft-landing to maintain tissue integrity. A two-layered matrix deposition method was used to improve mass spectral reproducibility across sections. Molecular images of the two subunits of alpha-crystallin and their modifications over approximately one-half of a single tissue section were reconstituted from mass spectral data sets acquired in 250-microm steps. Identification of protein truncation products and confirmation of phosphorylation distribution patterns were performed by reverse-phase liquid chromatography of soluble extracts from specific tissue regions followed by tandem mass spectrometry (LC/MS/MS). RESULTS: Distinct distribution patterns were observed for the two subunits of alpha-crystallin and their modified forms. alphaA-crystallin showed extensive truncation across whole sections, especially in the nuclei, whereas alphaB-crystallin was observed to be relatively stable. Both alphaA-crystallin and alphaB-crystallin displayed the highest level of phosphorylation in the middle cortex region, a finding confirmed by LC/MS/MS analysis of dissected regions. CONCLUSIONS: A new imaging technique has been successfully applied to molecularly characterize the spatial distribution of lens proteins and their modifications in lens sections. The different distributions of alpha-crystallin revealed in this study provide new leads in the investigation of underlying physiological significance of the modified forms of the two alpha-crystallin subunits.
Authors:
Jun Han; Kevin L Schey
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural    
Journal Detail:
Title:  Investigative ophthalmology & visual science     Volume:  47     ISSN:  0146-0404     ISO Abbreviation:  Invest. Ophthalmol. Vis. Sci.     Publication Date:  2006 Jul 
Date Detail:
Created Date:  2006-06-26     Completed Date:  2006-08-07     Revised Date:  2007-11-14    
Medline Journal Info:
Nlm Unique ID:  7703701     Medline TA:  Invest Ophthalmol Vis Sci     Country:  United States    
Other Details:
Languages:  eng     Pagination:  2990-6     Citation Subset:  IM    
Affiliation:
Department of Cell and Molecular Pharmacology, Medical University of South Carolina, Charleston, South Carolina 29425, USA.
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MeSH Terms
Descriptor/Qualifier:
Animals
Cattle
Chromatography, High Pressure Liquid
Lens, Crystalline / metabolism*
Phosphorylation
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / methods*
alpha-Crystallin A Chain / metabolism*
alpha-Crystallin B Chain / metabolism*
Grant Support
ID/Acronym/Agency:
EY13462/EY/NEI NIH HHS; R24 EY14793/EY/NEI NIH HHS
Chemical
Reg. No./Substance:
0/alpha-Crystallin A Chain; 0/alpha-Crystallin B Chain

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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