Synthesis and structure-activity study of myxoma virus growth factor.
Myxoma virus growth factor (MGF) is an 85-residue peptide derived from the gene product of a DNA tumor virus that infects rabbits. The carboxyl domain of MGF possesses about 40% sequence homology with the epidermal growth factor (EGF). This EGF-like domain covering residues 30-83 was synthesized and found to possess putative activities of EGF. It was, however, about 200-fold less active than EGF in the competitive binding of EGF receptor in A431 cells and the stimulation of [3H]-thymidine uptake in NRK 49F cells. MGF(30-83) is a basic and a hydrophobic peptide rich in beta-sheet structure. These features in MGF tend to promote aggregation, leading to precipitation even in strongly denaturing solutions. Thus, the refolding of MGF was achieved with difficulty and resulted in low yield. To increase the synthetic yield of MGF(30-83), a cluster of acidic amino acids was added to the NH2-terminus of MGF(30-83). This approach was found to be effective in minimizing the refolding difficulties and allowed accessibility to the synthesis of analogues in this class of compounds. The relationships of structure and function of MGF were studied by using analogues with point substitution by the corresponding D-amino acid or by Ala at position 44 or 52 and analogues with deletion of basic residues from the amino terminus. Modifications of both the receptor contact and the structural residues greatly reduced the potency of MGF(30-83), and the overall result correlated well with the known structure-activity of the EGF family.
Lin
Y Z
YZ
Rockefeller University, New York, New York 10021-6399.
Ke
X H
XH
Tam
J P
JP
D000595:Amino Acid Sequence; D002460:Cell Line; D004261:DNA Replication / Q000187:drug effects; D066246:ErbB Receptors / Q000187:drug effects / Q000378:metabolism; D006133:Growth Substances / Q000138:chemical synthesis* / Q000494:pharmacology; D006801:Humans; D007202:Indicators and Reagents; D036341:Intercellular Signaling Peptides and Proteins; D013058:Mass Spectrometry; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009233:Myxoma virus; D010455:Peptides / Q000138:chemical synthesis* / Q000494:pharmacology; D011487:Protein Conformation; D013329:Structure-Activity Relationship
D006133:Growth Substances; D007202:Indicators and Reagents; D036341:Intercellular Signaling Peptides and Proteins; D010455:Peptides; C068110:myxoma virus growth factor; D066246:ErbB Receptors