THE HUMAN KERATINOCYTE 2-DIMENSIONAL GEL PROTEIN DATABASE (UPDATE 1995) - MAPPING COMPONENTS OF SIGNAL-TRANSDUCTION PATHWAYS

The master two-dimensional (2-D) gel database of human keratinocytes c urrently lists 3154 cellular proteins (2224 isoelectric focusing, IEF; and 930 nonequilibrium pH gradient electrophoresis, NEPHGE), many of which correspond to post-translational modifications. 1082 polypeptide s have been identified (protein name, organelle components, etc.) usin g a procedure or a combination of procedures that include (i) comigrat ion with known human proteins, (ii) 2-D gel immunoblotting using speci fic antibodies, (iii) microsequencing of Coomassie Brilliant Blue stai ned proteins, (iv) mass spectrometry, (v) vaccinia virus expression of full length cDNAs, and (vi) in vitro transcription/translation of ful l-length cDNAs. This year, special emphasis has been given to the iden tification of signal transduction components by using 2-D gel immunobl otting of crude keratinocyte lysates in combination with enhanced chem oluminescence (ECL) detection. Identified proteins are listed both in alphabetical order and with increasing SSP number, together with their M(r), pI, cellular localization and credit to the investigator(s) tha t aided in the identification. Ultimately, the aim of the comprehensiv e database is to gather - through a systematic study of ekeratinocytes - qualitative and quantitative information on proteins and their gene s that may allow us to identify abnormal patterns of gene expression a nd to pinpoint signaling pathways and components affected in various s kin diseases, cancer included.