Chemical synthesis and spontaneous folding of a multidomain protein: anticoagulant microprotein S.
Because of recent high-yield native ligation techniques, chemical synthesis of larger multidomain bioactive proteins is rapidly coming within reach. Here we describe the total chemical synthesis of a designed "microprotein S," comprising the gamma-carboxyglutamic acid-rich module, the thrombin-sensitive module, and the first epidermal growth factor-like module of human plasma protein S (residues 1-116). Synthetic microprotein S expressed anticoagulant cofactor activity for activated protein C in the down-regulation of blood coagulation, and the anticoagulant activity of microprotein S was not neutralized by C4b-binding protein, a natural inhibitor of native protein S in plasma. The correct folding of this complex multidomain protein was enhanced compared with individual modules because the gamma-carboxyglutamic acid-rich module and the thrombin-sensitive module markedly facilitated correct folding of the first epidermal growth factor-like module compared with folding of the first epidermal growth factor-like module alone. These results demonstrate that total chemical synthesis of proteins offers an effective way to generate multidomain biologically active proteins.
Hackeng
T M
TM
Department of Molecular and Experimental Medicine, Skaggs Institute for Chemical Biology, La Jolla, CA 92037, USA. t.hackeng@bioch.unimaas.nl
Fernández
J A
JA
Dawson
P E
PE
Kent
S B
SB
Griffin
J H
JH
D015055:1-Carboxyglutamic Acid / Q000737:chemistry*; D000595:Amino Acid Sequence; D000925:Anticoagulants / Q000737:chemistry*; D004815:Epidermal Growth Factor / Q000737:chemistry*; D006801:Humans; D008969:Molecular Sequence Data; D017510:Protein Folding; D017293:Protein S / Q000737:chemistry*; D017434:Protein Structure, Tertiary
D000925:Anticoagulants; D017293:Protein S; D015055:1-Carboxyglutamic Acid; D004815:Epidermal Growth Factor