Peptide mapping on HIV polypeptides expressed in Escherichia coli. Quality control of different batches and identification of tryptic fragments containing residues of aromatic amino acids or cysteine.
Peptide mapping was used for the quality control of different batches of the recombinant HIV proteins p24 core and p24-gp41, expressed in Escherichia coli. These proteins comprise gag and env region polypeptides of the virus and may serve as suitable components in the diagnosis of HIV infections. The proteins were digested with trypsin and the mixtures were subjected to peptide mapping to prove batch equivalence of p24-gp41 and to isolate fragments of the p24-gp41 digest that differ from those of the p24 core digest. The proteins were reduced with dithiothreitol and the cysteine residues were derivatized by addition of 4-vinylpyridine. Peptide mapping was performed by means of reversed-phase high-performance liquid chromatography. Batch equivalence was proved by comparison of the maps. Peaks present in one map but not in the other were considered to be due to sequence differences or variability in digestion.
Renlund
S
S
Pharmacia LKB Biotechnology, Uppsala, Sweden.
Klintrot
I M
IM
Nunn
M
M
Schrimsher
J L
JL
Wernstedt
C
C
Hellman
U
U
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D003545:Cysteine / Q000032:analysis; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial; D015967:Gene Expression Regulation, Viral; D015683:Gene Products, gag / Q000737:chemistry / Q000235:genetics; D006678:HIV / Q000032:analysis* / Q000235:genetics; D016655:HIV Core Protein p24; D015700:HIV Envelope Protein gp41 / Q000737:chemistry / Q000235:genetics; D006868:Hydrolysis; D007202:Indicators and Reagents; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry; D010449:Peptide Mapping; D010455:Peptides / Q000737:chemistry* / Q000235:genetics; D011786:Quality Control; D013056:Spectrophotometry, Ultraviolet; D014357:Trypsin; D014758:Viral Core Proteins / Q000737:chemistry / Q000235:genetics
D000596:Amino Acids; D015683:Gene Products, gag; D016655:HIV Core Protein p24; D015700:HIV Envelope Protein gp41; D007202:Indicators and Reagents; D010446:Peptide Fragments; D010455:Peptides; D014758:Viral Core Proteins; D014357:Trypsin; D003545:Cysteine