Inhibitory peptides of the sulfotransferase domain of the heparan sulfate enzyme, N-deacetylase-N-sulfotransferase-1.
N-Deacetylase-N-sulfotransferase 1 (Ndst1) catalyzes the initial modification of heparan sulfate and heparin during their biosynthesis by removal of acetyl groups from subsets of N-acetylglucosamine units and subsequent sulfation of the resulting free amino groups. In this study, we used a phage display library to select peptides that interact with Ndst1, with the aim of finding inhibitors of the enzyme. The phage library consisted of cyclic random 10-mer peptides expressed in the phage capsid protein pIII. Selection was based on the ability of engineered phage to bind to recombinant murine Ndst1 (mNdst1) and displacement with heparin. Peptides that were enriched through multiple cycles of binding and disassociation displayed two specific sequences, CRGWRGEKIGNC and CNMQALSMPVTC. Both peptides inhibited mNdst1 activity in vitro, however, by distinct mechanisms. The peptide CRGWRGEKIGNC presents a chemokine-like repeat motif (BXX, where B represents a basic amino acid and X is a noncharged amino acid) and binds to heparan sulfate, thus blocking the binding of substrate to the enzyme. The peptide NMQALSMPVT inhibits mNdst1 activity by direct interaction with the enzyme near the active site. The discovery of inhibitory peptides in this way suggests a method for developing peptide inhibitors of heparan sulfate biosynthesis.
Gesteira
Tarsis F
TF
Departamentos de Bioquímica, Universidade Federal de Sa˜o Paulo, 04044-020 Sa˜o Paulo, Brazil.
Coulson-Thomas
Vivien J
VJ
Taunay-Rodrigues
Alessandro
A
Oliveira
Vitor
V
Thacker
Bryan E
BE
Juliano
Maria A
MA
Pasqualini
Renata
R
Arap
Wadih
W
Tersariol
Ivarne L S
IL
Nader
Helena B
HB
Esko
Jeffrey D
JD
Pinhal
Maria A S
MA
D020816:Amino Acid Motifs; D000818:Animals; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D004791:Enzyme Inhibitors / Q000737:chemistry*; D006497:Heparan Sulfate / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D006801:Humans; D051379:Mice; D019151:Peptide Library; D010456:Peptides, Cyclic / Q000737:chemistry* / Q000235:genetics; D017434:Protein Structure, Tertiary; D015238:Sulfotransferases / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000235:genetics / Q000378:metabolism
D004791:Enzyme Inhibitors; D006497:Heparan Sulfate; D019151:Peptide Library; D010456:Peptides, Cyclic; D015238:Sulfotransferases; C021350:heparitin sulfotransferase