Properties of Human Prolactin (PRL) and H27A-PRL, a Mutant That Does Not Bind Zn ++

Zhenyu Sun, P. Shirley Li, Priscilla S. Dannies, James Lee

Research output: Contribution to journalArticle

24 Citations (Scopus)

Abstract

The ability of protein hormones to self-associate is likely to play an important role in concentrating hormones into secretory granules; therefore, the aggregation properties of human PRL and H27A mutant were investigated. Human PRL bound 65Zn++; the Scatchard analysis was convex up, and limited by the solubility of PRL, but at least 0.7 mole Zn++ bound per mole of PRL. Binding of 65Zn++ to H27A-PRL was greatly reduced. The biological activity in an Nb2 cell assay and the circular dichroism spectrum of wild type and H27A-PRL were similar, indicating the H27A mutant folded correctly, and the binding of Zn++ to the high affinity site is not essential for biological activity. Dynamic light scattering measurements indicated 10 and 20 μM Zn++ caused some aggregation of both wild type and H27A-PRL. Sedimentation equilibrium analysis indicated that PRL is primarily a monomer in the absence of Zn++ and that there is increasing self-association in the presence of 5 and 10 μM Zn++. The mutant H27A exhibited a greater tendency to aggregate without changing detectably the mode of association. Although human PRL binds Zn++ as human GH does, it differs in that the ability to bind Zn++ and to self-associate were decoupled in PRL. Human PRL must have two types of interactions with Zn++; one is binding to a site involving histidine 27, and the other is weaker interactions that induce self-association of PRL.

Original languageEnglish
Pages (from-to)265-271
Number of pages7
JournalMolecular Endocrinology
Volume10
Issue number3
StatePublished - 1996

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Prolactin
Hormones
Secretory Vesicles
Circular Dichroism
Histidine
Solubility

ASJC Scopus subject areas

  • Molecular Biology
  • Endocrinology, Diabetes and Metabolism

Cite this

Sun, Z., Li, P. S., Dannies, P. S., & Lee, J. (1996). Properties of Human Prolactin (PRL) and H27A-PRL, a Mutant That Does Not Bind Zn ++. Molecular Endocrinology, 10(3), 265-271.

Properties of Human Prolactin (PRL) and H27A-PRL, a Mutant That Does Not Bind Zn ++. / Sun, Zhenyu; Li, P. Shirley; Dannies, Priscilla S.; Lee, James.

In: Molecular Endocrinology, Vol. 10, No. 3, 1996, p. 265-271.

Research output: Contribution to journalArticle

Sun, Z, Li, PS, Dannies, PS & Lee, J 1996, 'Properties of Human Prolactin (PRL) and H27A-PRL, a Mutant That Does Not Bind Zn ++', Molecular Endocrinology, vol. 10, no. 3, pp. 265-271.
Sun, Zhenyu ; Li, P. Shirley ; Dannies, Priscilla S. ; Lee, James. / Properties of Human Prolactin (PRL) and H27A-PRL, a Mutant That Does Not Bind Zn ++. In: Molecular Endocrinology. 1996 ; Vol. 10, No. 3. pp. 265-271.
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