Total chemical synthesis of human activin ?A[12-116] and related large-loop polypeptides

Abstract

We report here the synthesis, purification, and characterization of several large polypeptides related to the human activin βA subunit and their cyclic counterparts. In particular, we describe for the first time the total chemical synthesis of a 105‐mer polypeptide, des[1–11] activin βA, and related large‐loop polypeptide, by an optimized solid phase synthetic protocol based on 9‐flouroenylmethyoxycarbonyl (Fmoc) chemistry. These studies show that automated chemical synthesis utilizing Fmoc‐based solid phase synthetic strategies provides a practical alternative to recombinant DNA technology for the production of activin‐related subunits, with the opportunity to rapidly provide different analogues and structural variants for subsequent structure–function and associated biophysical investigations. © 2001 John Wiley & Sons, Inc. Biopolymers (Pept Sci) 60: 279–289, 2001

Thermodynamic analysis of the interaction between proteins and solid surfaces: application to liquid chromatography

“…(29). In the more general case of various a-, bor g-amino acid side chains each member with have a unique set of GMPs depending on the chemical nature of the interaction environment and this has lead to the tabulations of different hydrophobicity scales as described by Wilce et al 121 The relationships implicit to eqs (29)-(34), linking the free energy increments with the structural elements of a group of substances, underpins the remarkable power of chromatography to separate molecular species that differ only slightly in terms of a single structural feature, such as differences in a single amide group, substitution or deletion of an amino acid residue in large polypeptides, such as activin b A produced 160 by solid phase peptide synthesis or the resolution of polypeptide diastereomers or topomers. Moreover, when using quantitative chromatographic approaches it is of great importance to understand that the free energy difference for two compounds that differ in a structural element is proportional to the corresponding free energy change for that structural elements but not the rest of the molecule per se.…”