PMID- 1569054
OWN - NLM
STAT- MEDLINE
DCOM- 19920528
LR  - 20210210
IS  - 0021-9258 (Print)
IS  - 0021-9258 (Linking)
VI  - 267
IP  - 12
DP  - 1992 Apr 25
TI  - Purification and characterization of a functionally homogeneous 60-kDa species of 
      the retinoblastoma gene product.
PG  - 7971-4
AB  - The retinoblastoma susceptibility gene (RB) encodes a 928-amino acid protein 
      (pRB) that is hypothesized to function in a pathway that restricts cell 
      proliferation. The immortalizing proteins from three distinct DNA tumor viruses 
      (SV40 large T antigen, adenovirus E1a, and human papilloma virus Type 16 E7) have 
      been shown to interact with RB protein through two noncontiguous regions 
      comprised of amino acids 393-572 (domain A) and 646-772 (domain B). We 
      constructed a truncated form of RB (RB p60) that retains these two domains but 
      eliminates the N-terminal 386 amino acids of RB. RB p60 was expressed in 
      Escherichia coli in inclusion bodies. After solubilization, it was refolded in 
      the presence of magnesium chloride, and the active protein was isolated with an 
      E7 peptide affinity column. The protein that elutes from this column is 
      functionally homogenous in its ability to bind immobilized E7 protein. Thermal 
      denaturation studies provide additional evidence for the conformational 
      homogeneity of the isolated protein. This purification scheme allows the 
      isolation of significant amounts of RB p60 protein that is suitable for 
      structural and functional studies.
FAU - Edwards, G M
AU  - Edwards GM
AD  - Department of Cancer Research, Merck Sharp and Dohme Research Laboratories, West 
      Point, Pennsylvania 19486.
FAU - Huber, H E
AU  - Huber HE
FAU - DeFeo-Jones, D
AU  - DeFeo-Jones D
FAU - Vuocolo, G
AU  - Vuocolo G
FAU - Goodhart, P J
AU  - Goodhart PJ
FAU - Maigetter, R Z
AU  - Maigetter RZ
FAU - Sanyal, G
AU  - Sanyal G
FAU - Oliff, A
AU  - Oliff A
FAU - Heimbrook, D C
AU  - Heimbrook DC
LA  - eng
PT  - Journal Article
PL  - United States
TA  - J Biol Chem
JT  - The Journal of biological chemistry
JID - 2985121R
RN  - 0 (Oncogene Proteins, Viral)
RN  - 0 (Papillomavirus E7 Proteins)
RN  - 0 (Retinoblastoma Protein)
RN  - 0 (oncogene protein E7, Human papillomavirus type 16)
RN  - 9007-49-2 (DNA)
SB  - IM
EIN - J Biol Chem 1992 Jul 5;267(19):13780
MH  - Chromatography, Affinity
MH  - Circular Dichroism
MH  - DNA/genetics
MH  - Electrophoresis, Polyacrylamide Gel
MH  - Enzyme-Linked Immunosorbent Assay
MH  - Escherichia coli/genetics
MH  - Gene Expression
MH  - Genes, Bacterial
MH  - Hot Temperature
MH  - Humans
MH  - Oncogene Proteins, Viral/metabolism
MH  - Papillomavirus E7 Proteins
MH  - Plasmids
MH  - Protein Conformation
MH  - Protein Denaturation
MH  - Retinoblastoma Protein/*isolation & purification/metabolism
EDAT- 1992/04/25 00:00
MHDA- 1992/04/25 00:01
CRDT- 1992/04/25 00:00
PHST- 1992/04/25 00:00 [pubmed]
PHST- 1992/04/25 00:01 [medline]
PHST- 1992/04/25 00:00 [entrez]
AID - S0021-9258(18)42393-9 [pii]
PST - ppublish
SO  - J Biol Chem. 1992 Apr 25;267(12):7971-4.