Hypusine-containing proteins and their modifying enzymes

Thesis/Dissertation · Fri Jan 01 04:00:00 EST 1988

OSTI ID:6948677

Dou, Q

The aim of this study is to purify and characterize this protein and to investigate general properties of its modifying enzymes. I developed a simple four-step procedure for purifying the hyp-18K protein. This procedure resulted in an 800-fold purification of the radio-labeled protein from NB-15 mouse neuroblastoma cells. Two hypusine-containing proteins with an apparent molecular weight of 18 kDa and 20 kDa were also identified by metabolic labeling in chick embryo fibroblasts. The hyp-18K in both NB cells and chick embryo fibroblastes exhibited two isoforms: acidic form (hyp-18K{sup a}) and basic form (hyp-18K{sup b}). I also purified both hyp-20K and hyp-18K isoforms from chick embryos to homogeneity. V8 protease peptide map analysis indicated that hyp-18K{sup b} isolated from chick embryos was identical, while hyp-18K{sup a} and hyp-20K were similar, to elF-4D. The chick embryo hyp-18K{sup b}, but not hyp-18K{sup a}, co-migrated with elF-4D. When NB cell lysates were incubated with ({sup 3}H)spermidine at pH 9.5, only hyp-18K{sup b} was labeled. Lysates, labeled at pH 9.5 and then adjusted to pH 7.2, gave both hyp-18K{sup a} and hyp-18K{sup b}. These data provided evidence that hyp-18K{sup a} is derived from hyp-18K{sup b}.

• Research Organization: Rutgers--the State Univ., New Brunswick, NJ (USA)
• OSTI ID: 6948677
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
AMINES
ANIMAL CELLS
ANIMALS
BIRDS
CHEMICAL COMPOSITION
CHICKENS
EMBRYOS
ENZYMES
FOWL
FRACTIONATION
HYDROGEN COMPOUNDS
HYDROLASES
ISOTOPE APPLICATIONS
MAMMALS
MICE
MOLECULAR WEIGHT
ORGANIC COMPOUNDS
PEPTIDE HYDROLASES
PROTEINS
RODENTS
SEPARATION PROCESSES
SPERMIDINE
TRACER TECHNIQUES
TRITIUM COMPOUNDS
TUMOR CELLS
VERTEBRATES