PMID:
1286667
Authors:
Rasmussen HH, van Damme J, Puype M, Gesser B, Celis JE, Vandekerckhove J.
Title:
Microsequences of 145 proteins recorded in the two-dimensional gel protein
database of normal human epidermal keratinocytes.
Journal:
Electrophoresis. 1992 Dec;13(12):960-9. doi: 10.1002/elps.11501301199.
Abstract:
Microsequencing of proteins recovered from two-dimensional (2-D) gels is being
used systematically to identify proteins in the master human keratinocyte 2-D gel
database. To date, about 250 protein spots recorded in human 2-D gel databases
have been microsequenced and, of these, 145 are recorded in the keratinocyte
database under the entry partial amino acid sequence. Coomassie Brilliant
Blue-stained protein spots cut from several (up to 40) dry gels were concentrated
by elution-concentration gel electrophoresis, electroblotted onto PVDF membranes
and digested in situ with trypsin. Eluting peptides were separated by
reversed-phase HPLC, collected individually and sequenced. Computer search using
the FASTA and TFASTA programs from Genetics Computer Group indicated that 110 of
the microsequenced polypeptides shared significant similarity with proteins
contained in the PIR, Mipsx or GenEMBL databases. Only 35 polypeptides
corresponded to hitherto unknown proteins. Peptide sequences of all 145 proteins
are listed together with their coordinates (apparent molecular weight and pI) in
the keratinocyte database.
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