Sequence simplification and the intra- and intermolecular self-recognition properties of vasopressin/neurophysin biosynthetic precursor.
The self-assembly properties of the arginine 8-vasopressin/bovine neurophysin II (AVP/BNPII) biosynthetic precursor were studied using glycopeptide-deleted and sequence-redesigned semisynthetic derivatives. Semisynthetic precursors were prepared by chemically coupling synthetic vasopressinyl sequence domains and native protein-derived neurophysin II domain. Measurement of precursor-protein association by the extent of affinity chromatographic retardation on agarose-immobilized BNPII verified that the semisynthetic precursor with native AVP sequence has an enhanced self-association propensity similar to that predicted for native precursor. Here, the stabilizing contacts between hormone and neurophysin domains, mainly the positively charged protonated alpha-amino group and tyrosyl 2 side chain of the hormone, are retained. Semisynthetic precursor variants in which the hormone domain is sequence-simplified by introducing alanyl residues in positions not considered important for neurophysin recognition show non-reduced association to BNPII. In contrast, removal of one of the main contact elements between hormone and neurophysin by acetylation of the hormone alpha-amino group abolishes potentiation of precursor self-association. The results show that the presence of the C-terminal glycopeptide sequence domain of native vasopressin precursor is not required to promote self-assembly of the precursor. The data verify the view proposed for the oxytocinyl precursor that intramolecular domain interaction is the triggering event which promotes the increase in affinity of precursor self-association (intermolecular self-recognition). The data also define some of the intramolecular self-recognition elements in the folded precursor required for the high affinity intermolecular self-recognition.
Fassina
G
G
National Institute of Diabetes and Digestive and Kidney Diseases, Bethesda, MD 20892.
Chaiken
I M
IM
D000595:Amino Acid Sequence; D000818:Animals; D001127:Arginine Vasopressin / Q000235:genetics / Q000378:metabolism*; D002417:Cattle; D055598:Chemical Phenomena; D002621:Chemistry; D002846:Chromatography, Affinity; D003488:Cyanogen Bromide; D014644:Genetic Variation; D006023:Glycoproteins / Q000302:isolation & purification; D007700:Kinetics; D008954:Models, Biological; D008969:Molecular Sequence Data; D009481:Neurophysins / Q000378:metabolism*; D010902:Pituitary Gland / Q000032:analysis; D011498:Protein Precursors / Q000235:genetics / Q000378:metabolism*
D006023:Glycoproteins; D009481:Neurophysins; D011498:Protein Precursors; D001127:Arginine Vasopressin; D003488:Cyanogen Bromide