Novel ATP-binding heat-inducible protein of Mr = 37,000 that is sensitive to transformation in BALB/3T3 cells

Journal Article · Fri Jun 01 00:00:00 EDT 1990 · Journal of Cellular Physiology; (USA)

DOI:https://doi.org/10.1002/jcp.1041430324· OSTI ID:6643896

Nakai, A; Hirayama, C; Ohtsuka, K; Hirayoshi, K; Nagata, K ^[1]

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1. Kyoto Univ. (Japan)

Using affinity chromatography on ATP-agarose, we have identified a major ATP-binding protein in Nonidet P-40 extracts of avian and mammalian cells labeled with (35S)methionine. After washing ATP-agarose beads with high-ionic-strength buffer (0.4 M NaCl), the 37-kD protein was shown to be one of the major ATP-binding proteins while p72 and grp78, which are members of the hsp70 family, also bound to ATP-agarose. This protein consisted of several spots on two-dimensional gel electrophoresis. The isoelectric point of the most basic spot was approximately 9.2 in chick embryo fibroblasts, whereas it was about 8.8 in mouse 3T3 cells. The identities of these proteins in mouse and chick cells were confirmed by peptide mapping. After heat-shock treatment of BALB/3T3 cells, the major heat-shock protein, hsp70, was shown to be induced very rapidly after heat shock and was recovered in the ATP-binding fraction. Besides hsp70, a 37-kD protein was also found to be induced by heat shock. This protein was drastically induced by treating the cells with alpha,alpha'-dipyridyl, an iron chelating reagent, but not with sodium arsenite, calcium ionophore, or tunicamycin. The synthesis and the total amount of this ATP-binding protein increased in mouse 3T3 cells transformed by simian virus 40, methylcholanthrene, or activated c-Ha-ras oncogene compared to their normal counterparts. The incorporation of (32P)orthophosphate was not detected in either normal or transformed cells. These studies established that a major ATP-binding protein of Mr = 37,000 is a heat-inducible protein and that the synthesis of this protein is regulated by malignant transformation.

• OSTI ID: 6643896
• Journal Information: Journal of Cellular Physiology; (USA), Vol. 143:3; ISSN 0021-9541
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
3-METHYLCHOLANTHRENE
BIOLOGICAL EFFECTS
ATP
BIOCHEMICAL REACTION KINETICS
FIBROBLASTS
ONCOGENIC TRANSFORMATIONS
PROTEINS
AMINO ACID SEQUENCE
ARSENIC
BIOLOGICAL SHOCK
CELL CULTURES
CHICKENS
ELECTROPHORESIS
EMBRYOS
HEAT
LABELLING
METHIONINE
MICE
PHOSPHORUS 32
SULFUR 35
AMINO ACIDS
ANIMAL CELLS
ANIMALS
AROMATICS
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BIRDS
CARBOXYLIC ACIDS
CELL TRANSFORMATIONS
CONNECTIVE TISSUE CELLS
DAYS LIVING RADIOISOTOPES
DRUGS
ELEMENTS
ENERGY
EVEN-ODD NUCLEI
FOWL
HYDROCARBONS
ISOTOPES
KINETICS
LIGHT NUCLEI
LIPOTROPIC FACTORS
MAMMALS
MOLECULAR STRUCTURE
NUCLEI
NUCLEOTIDES
ODD-ODD NUCLEI
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC SULFUR COMPOUNDS
PATHOLOGICAL CHANGES
PHOSPHORUS ISOTOPES
POLYCYCLIC AROMATIC HYDROCARBONS
RADIOISOTOPES
REACTION KINETICS
RODENTS
SEMIMETALS
SOMATIC CELLS
SULFUR ISOTOPES
VERTEBRATES
550201* - Biochemistry- Tracer Techniques