The primary structure of the lymphocyte pore-forming protein perforin: partial amino acid sequencing and determination of isoelectric point.
The murine lymphocyte pore-forming protein (PFP) was purified to apparent homogeneity by successive steps of liquid chromatography. Monospecific antibodies were raised against purified PFP that detect only one protein band in murine CTL lines. 25% of the primary sequence of PFP (134 amino acids) was determined by amino terminal analysis of the purified protein and of some of its enzymatic cleavage products. These primary sequences were identical to sequences deduced by cDNA cloning. By isoelectric focusing, PFP was found to have a pI of 6.4. On the chromatofocusing column Mono P, however, PFP was found to elute at pH 4.7. This suggests a tertiary structure for monomeric PFP that is enriched in surface acidic amino acids.
Persechini
P M
PM
Laboratory of Cellular Physiology and Immunology, Rockefeller University, New York, N.Y. 10021.
Young
J D
JD
D000595:Amino Acid Sequence; D000818:Animals; D002118:Calcium / Q000494:pharmacology; D002460:Cell Line; D002845:Chromatography; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D015151:Immunoblotting; D007525:Isoelectric Focusing; D007526:Isoelectric Point; D008562:Membrane Glycoproteins; D008565:Membrane Proteins / Q000302:isolation & purification; D051379:Mice; D008969:Molecular Sequence Data; D054353:Perforin; D052899:Pore Forming Cytotoxic Proteins
D008562:Membrane Glycoproteins; D008565:Membrane Proteins; D052899:Pore Forming Cytotoxic Proteins; D054353:Perforin; D004247:DNA; D002118:Calcium