Engineered production of cancer targeting peptide (CTP)-containing C-1027 in Streptomyces globisporus and biological evaluation.
Conjugation of cancer targeting peptides (CTPs) with small molecular therapeutics has emerged as a promising strategy to deliver potent (but typically nonspecific) cytotoxic agents selectively to cancer cells. Here we report the engineered production of a CTP (NGR)-containing C-1027 and evaluation of its activity against selected cancer cell lines. C-1027 is an enediyne chromoprotein produced by Streptomyces globisporus, consisting of an apo-protein (CagA) and an enediyne chromophore (C-1027). NGR is a CTP that targets CD13 in tumor vasculature. S. globisporus SB1026, a recombinant strain engineered to encode CagA with the NGR sequence fused at its C-terminus, directly produces the NGR-containing C-1027 that is equally active as the native C-1027. Our results demonstrate the feasibility to produce CTP-containing enediyne chromoproteins by metabolic pathway engineering and microbial fermentation and will inspire efforts to engineer other CTP-containing drug binding proteins for targeted delivery.
Division of Pharmaceutical Sciences, University of Wisconsin, Madison, WI 53705, USA.
Wenli
W
Li
Department of Cancer Biology, The Scripps Research Institute, Jupiter, FL 33458, USA.
Xiuling
X
Li
Department of Chemistry, The Scripps Research Institute, Jupiter, FL 33458, USA.
Tingting
T
Huang
Department of Chemistry, The Scripps Research Institute, Jupiter, FL 33458, USA.
Qihui
Q
Teng
Department of Chemistry, The Scripps Research Institute, Jupiter, FL 33458, USA.
Ivana
I
Crnovcic
Department of Cancer Biology, The Scripps Research Institute, Jupiter, FL 33458, USA; Department of Molecular Therapeutics, The Scripps Research Institute, Jupiter, FL 33458, USA.
Christoph
C
Rader
Division of Pharmaceutical Sciences, University of Wisconsin, Madison, WI 53705, USA; Department of Chemistry, The Scripps Research Institute, Jupiter, FL 33458, USA; Department of Molecular Therapeutics, The Scripps Research Institute, Jupiter, FL 33458, USA; Natural Products Library Initiative at The Scripps Research Institute, The Scripps Research Institute, Jupiter, FL 33458, USA. Electronic address: shenb@scripps.edu.
Ben
B
Shen
D000617:Aminoglycosides / Q000096:biosynthesis / Q000494:pharmacology*; D000970:Antineoplastic Agents / Q000494:pharmacology*; D001426:Bacterial Proteins / Q000096:biosynthesis / Q000494:pharmacology*; D045744:Cell Line, Tumor; D053281:Enediynes / Q000494:pharmacology*; D005240:Feasibility Studies; D005285:Fermentation; D006801:Humans; D015202:Protein Engineering; D013302:Streptomyces / Q000378:metabolism
D000617:Aminoglycosides; D000970:Antineoplastic Agents; D001426:Bacterial Proteins; D053281:Enediynes; C058024:C 1027
C-1027; Cancer; Cancer targeting peptide (CTP); Enediyne; Streptomyces globisporus