Expression of the hAng gene in Escherichia coli; isolation and characterization of human recombinant Ser-(-1) angiogenin.
Recombinant human angiogenin has been synthesized in Escherichia coli with the aid of a human angiogenin gene (hAng) cloned by Neznanov et al (1990) from a human complementary DNA (cDNA) library. The gene has been expressed by use of a new type of expression vector called a 'TGATG vector' (plasmid pPR-TGATG-1; Mashko et al 1990a). The highest level of accumulation of the recombinant angiogenin (6%-8% of the total cell protein) was observed in E. coli strain BL21 carrying a temperature-amplifiable version of the plasmid. The synthesized polypeptide carries an additional serine residue at its N terminus in comparison with natural angiogenin. Furthermore, the initiator methionine residue of the recombinant protein is removed with high efficiency by E. coli terminal aminopeptidase. Simple procedures for purification of the recombinant angiogenin from the insoluble fraction of cell protein, and for refolding the protein allowed the isolation of almost 5 mg recombinant angiogenin g-1 wet bacterial biomass. The recombinant Ser-(-1) angiogenin displayed the same biological properties (specific RNAase activity and the ability to induce blood vessel growth on the sclera of experimental animals) as its natural counterpart isolated from human blood.
Lapidus
A L
AL
All-Union Research Institute of Genetics, Ministry of Medical Industry of the USSR, Moscow.
Mochul'skii
A V
AV
Podkovyrov
S M
SM
Lebedeva
M I
MI
Antipin
A A
AA
Izotova
L S
LS
Zagnit'ko
O P
OP
Komolova
G S
GS
Klesov
A A
AA
Veiko
V P
VP
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002196:Capillaries / Q000187:drug effects; D004926:Escherichia coli / Q000235:genetics*; D005260:Female; D005822:Genetic Vectors; D006801:Humans; D008297:Male; D008969:Molecular Sequence Data; D009363:Neoplasm Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000502:physiology; D010957:Plasmids / Q000235:genetics; D014176:Protein Biosynthesis; D011506:Proteins / Q000235:genetics / Q000502:physiology; D051381:Rats; D011994:Recombinant Proteins; D012259:Ribonuclease, Pancreatic; D012260:Ribonucleases / Q000096:biosynthesis / Q000187:drug effects; D012590:Sclera / Q000098:blood supply; D014162:Transfection
D009363:Neoplasm Proteins; D011506:Proteins; D011994:Recombinant Proteins; D012260:Ribonucleases; C046366:angiogenin; D012259:Ribonuclease, Pancreatic