Characterisation of low abundance wool proteins through novel differential extraction techniques.
Fibres from human hair and wool are characterised by two main types of proteins: intermediate filament proteins (IFPs) and keratin associated proteins (KAPs). The IFPs, comprising over 50% of the fibre, tend to dominate 2-D electrophoretic maps, hindering identification of the less-abundant KAPs. This has been compounded in wool fibres by the relatively limited amount of sequence information available, with approximately 35 distinct protein sequences from ten KAP families being available, in contrast to human hair, where the sequences from well over 80 proteins from 26 KAP families are known. Additional complications include the high degree of homology within these families, ranging from 70 to 95%, and the dominance of cysteine residues in a number of KAP families with their high propensity to form cross-links. The lack of sequence information for wool KAPs has been partly overcome through the recent acquisition of new sequences. Fractionation of the proteins on the basis of their solubility with pH, urea and DTT concentration has resulted in protein extracts in which the IFP concentration has been considerably reduced. These improvements have enabled the identification of low-abundance proteins in 2-D electrophoretic maps and represent a significant advance in our knowledge of the wool proteome.
Plowman
Jeffrey E
JE
AgResearch, Christchurch, New Zealand. jeff.plowman@agresearch.co.nz
Deb-Choudhury
Santanu
S
Thomas
Ancy
A
Clerens
Stefan
S
Cornellison
Charisa D
CD
Grosvenor
Anita J
AJ
Dyer
Jolon M
JM
D000595:Amino Acid Sequence; D000818:Animals; D004229:Dithiothreitol / Q000737:chemistry; D004586:Electrophoresis / Q000379:methods*; D015180:Electrophoresis, Gel, Two-Dimensional; D006197:Hair / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D053564:Keratins, Hair-Specific / Q000737:chemistry / Q000302:isolation & purification; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010455:Peptides / Q000302:isolation & purification; D012756:Sheep; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods; D014508:Urea / Q000737:chemistry; D014935:Wool / Q000737:chemistry*
D053564:Keratins, Hair-Specific; D010455:Peptides; D014508:Urea; D004229:Dithiothreitol