Eye lens proteomics: from global approach to detailed information about phakinin and gamma E and F crystallin genes.
Exploration of the lenticular proteome poses a challenging and worthwhile undertaking as cataracts, the products of a disease phenotype elicited by this proteome, remains the leading cause of vision impairment worldwide. The complete ten day old lens proteome of Mus musculus C57BL/6J was resolved into 900 distinct spots by large gel carrier ampholyte based 2-DE. The predicted amino acid sequences of all 16 crystallins ubiquitous in mammals were corroborated by mass spectrometry (MS). In detailed individual spot analyses, the primary structure of the full murine C57BL/6J beaded filament component phakinin CP49 was sequenced by liquid chromatography/electrospray ionization-tandem MS and amended at two positions. This definitive polypeptide sequence was aligned to the mouse genome, thus identifying the entire C57BL/6J genomic coding region. Also, two murine C57/6J polypeptides, both previously classified as gamma F crystallin, were clearly distinguished by MS and electrophoretic mobility. Both were assigned to their respective genes, one of the polypeptides was reclassified as C57BL/6J gamma E crystallin. Building on these data and previous investigations an updated crystallin reference map was put forth and several non crystallin lenticular components were examined. These results represent the first part of a comprehensive investigation of the mouse lens proteome (http://www.mpiib-berlin.mpg.de/2D-PAGE) with emphasis on understanding genetic effects on proteins and disease development.
Hoehenwarter
Wolfgang
W
Max-Planck-Institut für Infektionsbiologie, 10117 Berlin, Germany.
Kumar
Nalin M
NM
Wacker
Maik
M
Zimny-Arndt
Ursula
U
Klose
Joachim
J
Jungblut
Peter R
PR
D000595:Amino Acid Sequence; D000818:Animals; D002853:Chromatography, Liquid; D003459:Crystallins / Q000235:genetics / Q000378:metabolism*; D015180:Electrophoresis, Gel, Two-Dimensional; D005136:Eye Proteins / Q000235:genetics / Q000378:metabolism*; D007381:Intermediate Filament Proteins / Q000235:genetics / Q000378:metabolism*; D007908:Lens, Crystalline / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D010449:Peptide Mapping; D040901:Proteomics; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D038222:gamma-Crystallins
D003459:Crystallins; D005136:Eye Proteins; D007381:Intermediate Filament Proteins; C107256:crystallin gammaE; D038222:gamma-Crystallins; C084808:phakinin