Polyamines metabolically labeled two cellular proteins in fibroblasts isolated from chick embryos
A hypusine-containing protein (Mw=18,000) has been reported to be present in all mammalian cells examined. The formation of the unusual amino acid residue in this 18,000-dalton protein is due to a posttranslational modification of lysine residue by spermidine. To search for an abundant source for the purification of this protein, they have examined possible existence of this protein in chick embryos using metabolical labeling method. Metabolical labeling of chick embryo fibroblasts prepared from the Day 11 embryos by (2,3-TH)putrescine resulted in two prominently labeled protein bands as shown by SDS-PAGE and fluorography. The apparent molecular weights of the labeled proteins were 20,000- and 18,000-daltons. Two-dimensional gel analysis indicated that the 20,000-dalton protein had a pI of 5.5 and the 18,000-dalton protein exhibited isoform structures with pI values ranging from 4.6 to 5.1. Peptide map analysis showed that the 18,000-dalton protein from chick embryos was identical to the 18,000-dalton protein isolated from mouse neuroblastoma cells. The purification procedure that they have developed for mouse neuroblastoma 18,000-dalton protein was found to be also applicable in isolating the 18,000-dalton protein from chick embryos. Both the 18,000- and the 20,000-dalton proteins in chick embryos were enriched after Cibacron-Blue column and omega-diaminooctyl-agarose column chromatography.
- Research Organization:
- Rutgers, The State Univ. of New Jersey, Piscataway
- OSTI ID:
- 6285911
- Report Number(s):
- CONF-870644-
- Journal Information:
- Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States), Journal Name: Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States) Vol. 46:6; ISSN FEPRA
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
59 BASIC BIOLOGICAL SCIENCES
AMINES
ANIMAL CELLS
ANIMALS
BIRDS
CHICKENS
CHROMATOGRAPHY
CONNECTIVE TISSUE CELLS
ELECTROPHORESIS
EMBRYOS
FIBROBLASTS
FOWL
FRACTIONATION
GENETIC MAPPING
ISOTOPE APPLICATIONS
LABELLED COMPOUNDS
LABELLING
MAPPING
MOLECULAR WEIGHT
ORGANIC COMPOUNDS
PROTEINS
PUTRESCINE
SEPARATION PROCESSES
SOMATIC CELLS
SPERMIDINE
TRACER TECHNIQUES
TRITIUM COMPOUNDS
VERTEBRATES