Protein-electroblotting and -microsequencing strategies in generating protein data bases from two-dimensional gels.
- 1 October 1989
- journal article
- research article
- Published by Proceedings of the National Academy of Sciences in Proceedings of the National Academy of Sciences
- Vol. 86 (20) , 7701-7705
- https://doi.org/10.1073/pnas.86.20.7701
Abstract
Coomassie blue-stained, heat-dried, and computer-imaged two-dimensional gels used to develop comprehensive human protein data bases served as the protein source to generate partial amino acid sequences. The protein spots were collected from multiple gels, rehydrated, concentrated by stacking into a new gel, electoblotted onto inert membranes, and in situ-digested with trypsin. Peptides eluting from the membranes were separated by HPLC and sequenced. Using this procedure, it was possible to generate partial sequences from 13 human proteins recorded in the amnion cell protein data base. Eight of these sequences matched those of proteins stored in data bases, demonstrating that a systematic analysis of proteins by computerized two-dimensional gel electrophoresis can be directly linked to protein microsequencing methods. The latter technique offers a unique opportunity to link information contained in protein data bases derived from the analysis of two-dimensional gels with forthcoming DNA sequence data on the human genome.This publication has 38 references indexed in Scilit:
- The QUEST System for Quantitative Analysis of Two-dimensional GelsJournal of Biological Chemistry, 1989
- Five distinct calcium and phospholipid binding proteins share homology with lipocortin I.Journal of Biological Chemistry, 1988
- Internal amino acid sequence analysis of proteins separated by one- or two-dimensional gel electrophoresis after in situ protease digestion on nitrocellulose.Proceedings of the National Academy of Sciences, 1987
- Primary structure of human nuclear ribonucleoprotein particle C proteins: conservation of sequence and domain structures in heterogeneous nuclear RNA, mRNA, and pre-rRNA-binding proteins.Molecular and Cellular Biology, 1987
- Amino acid sequence of a chicken heat shock protein derived from the complementary DNA nucleotide sequenceBiochemistry, 1986
- Purification and N-terminal amino acid sequence of proliferating cell nuclear antigen (PCNA)/cyclin and development of ELISA for anti-PCNA antibodies.The Journal of Immunology, 1985
- Amino acid sequence of equine platelet tropomyosinFEBS Letters, 1983
- [17] Isolation of microgram quantities of proteins from polyacrylamide gels for amino acid sequence analysisPublished by Elsevier ,1983
- Radioiodination of proteins in single polyacrylamide gel slices. Tryptic peptide analysis of all the major members of complex multicomponent systems using microgram quantities of total protein.Journal of Biological Chemistry, 1977
- Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4Nature, 1970