Recombinant hemoglobin A produced in transgenic swine: structural equivalence with human hemoglobin A.
Recombinant human hemoglobin A produced by coexpressing human alpha and beta globin genes in swine, and purified from the lysate of transgenic swine has been subjected to detailed protein chemical analysis. These structural studies involving laser desorption mass spectrometry, separation of globin chains by RPHPLC, amino terminal sequence analysis of the isolated globin chains, the tryptic peptide mapping of the purified globin chains and the amino acid composition analysis of the purified tryptic peptides of globin chains have established the primary structural equivalence of the globin chains of the transgenic swine derived hemoglobin A with that of human hemoglobin A. These results demonstrate that the transgenic swine system correctly translates the human alpha and beta globin m-RNA; carries out the correct cotranslational processing of globin chains, and does not introduce any unwanted post translational modifications into the mature chains. Thus, the transgenic swine expression system is an excellent approach for the production of HbA for developing an effective hemoglobin based oxygen carrier.
Rao
M J
MJ
Division of Hematology, Albert Einstein College of Medicine, Bronx, NY 10461.
Schneider
K
K
Chait
B T
BT
Chao
T L
TL
Keller
H
H
Anderson
S
S
Manjula
B N
BN
Kumar
R
R
Acharya
A S
AS
D000595:Amino Acid Sequence; D000818:Animals; D030801:Animals, Genetically Modified; D001802:Blood Substitutes / Q000737:chemistry / Q000302:isolation & purification; D005914:Globins / Q000235:genetics; D006441:Hemoglobin A / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics*; D006801:Humans; D015394:Molecular Structure; D008970:Molecular Weight; D010100:Oxygen / Q000378:metabolism; D010449:Peptide Mapping; D014176:Protein Biosynthesis; D011487:Protein Conformation; D011499:Protein Processing, Post-Translational; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D013552:Swine
D001802:Blood Substitutes; D011994:Recombinant Proteins; D005914:Globins; D006441:Hemoglobin A; D010100:Oxygen