scholarly journals From Consensus Sequence Peptide to High Affinity Ligand, a “Library Scan” Strategy

2001 ◽  
Vol 276 (15) ◽  
pp. 12235-12240 ◽  
Author(s):  
Ren-Hwa Yeh ◽  
Tae Ryong Lee ◽  
David S. Lawrence
Keyword(s):  
Consensus Sequence ◽  
Sh2 Domain ◽  
Amino Acid Sequences ◽  
Parallel Synthesis ◽  
Intact Protein ◽  
High Affinity ◽  
Affinity Ligand ◽  
Combinatorial Peptide Libraries ◽  
Synthesis Strategy ◽  
Consensus Peptide

A wide variety of proteins have been shown to recognize and bind to specific amino acid sequences on other proteins. These sequences can be readily identified using combinatorial peptide libraries. However, peptides containing these preferred sequences (“consensus sequence peptides”) typically display only modest affinities for the consensus sequence-binding site on the intact protein. In this report, we describe a parallel synthesis strategy that transforms consensus sequence peptides into high affinity ligands. The work described herein has focused on the Lck SH2 domain, which binds the consensus peptide acetyl-Tyr(P)-Glu-Glu-Ile-amide with aKDof 1.3 μm. We employed a strategy that creates a series of spatially focused libraries that challenge specific subsites on the target protein with a diverse array of functionality. The final lead compound identified in this study displayed a 3300-fold higher affinity for the Lck SH2 domain than the starting consensus sequence peptide.

Characterization of Glycoprotein Fraction from Carp Pituitaries Isolated Using Concanavalin A as the Affinity Ligand

1998 ◽  
Vol 63 (3) ◽  
pp. 434-440 ◽  
Author(s):  
Irena Hulová ◽  
Jana Barthová ◽  
Helena Ryšlavá ◽  
Václav Kašička
Keyword(s):  
Concanavalin A ◽  
Reversed Phase ◽  
Amino Acid Sequences ◽  
Gel Chromatography ◽  
Affinity Ligand ◽  
Sds Page ◽  
Terminal Amino ◽  
Α And Β Subunits

Glycoproteins that have affinity to Concanavalin A were isolated from the acetone-dried pituitaries of common carp (Cyprinus carpio L.). Two fractions of glycoproteins were separated using gel chromatography on Superdex 75HR. The fraction with lower molecular weight (30 000) corresponding to the carp gonadotropin cGtH II was composed of two subunits as determined using SDS-PAGE. This protein fraction was further divided into four components using reversed-phase HPLC. Two fractions were pure α and β subunits of cGtH II as follows from immunodetection and from determination of N-terminal amino acid sequences. The other two were a mixture of α and β subunits as was also revealed by N-terminal analysis. Capillary electrophoresis was also used for characterization of isolated glycoproteins.

scholarly journals Enhanced Arsenic Accumulation in Engineered Bacterial Cells Expressing ArsR

2004 ◽  
Vol 70 (8) ◽  
pp. 4582-4587 ◽  
Author(s):  
Jan Kostal ◽  
Rosanna Yang ◽  
Cindy H. Wu ◽  
Ashok Mulchandani ◽  
Wilfred Chen
Keyword(s):  
Cell Growth ◽  
Environmental Protection Agency ◽  
Arsenic Removal ◽  
Fusion Partner ◽  
Resting Cells ◽  
Bacterial Cells ◽  
High Affinity ◽  
Affinity Ligand ◽  
Arsenic Accumulation ◽  
Severe Reduction

ABSTRACT The metalloregulatory protein ArsR, which offers high affinity and selectivity toward arsenite, was overexpressed in Escherichia coli in an attempt to increase the bioaccumulation of arsenic. Overproduction of ArsR resulted in elevated levels of arsenite bioaccumulation but also a severe reduction in cell growth. Incorporation of an elastin-like polypeptide as the fusion partner to ArsR (ELP153AR) improved cell growth by twofold without compromising the ability to accumulate arsenite. Resting cells overexpressing ELP153AR accumulated 5- and 60-fold-higher levels of arsenate and arsenite than control cells without ArsR overexpression. Conversely, no significant improvement in Cd2+ or Zn2+ accumulation was observed, validating the specificity of ArsR. The high affinity of ArsR allowed 100% removal of 50 ppb of arsenite from contaminated water with these engineered cells, providing a technology useful to comply with the newly approved U.S. Environmental Protection Agency limit of 10 ppb. These results open up the possibility of using cells overexpressing ArsR as an inexpensive, high-affinity ligand for arsenic removal from contaminated drinking and ground water.

scholarly journals A novel radioiodinated high affinity ligand for the D2 -dopamine receptor

FEBS Letters ◽  
1984 ◽  
Vol 176 (2) ◽  
pp. 436-440 ◽  
Author(s):  
Nourdine Amlaiky ◽  
Brian F. Kilpatrick ◽  
Marc G. Caron
Keyword(s):  
Dopamine Receptor ◽  
D2 Dopamine Receptor ◽  
High Affinity ◽  
Affinity Ligand ◽  
High Affinity Ligand

scholarly journals High-Affinity Ligand Probes of CD22 Overcome the Threshold Set bycisLigands to Allow for Binding, Endocytosis, and Killing of B Cells

2006 ◽  
Vol 177 (5) ◽  
pp. 2994-3003 ◽  
Author(s):  
Brian E. Collins ◽  
Ola Blixt ◽  
Shoufa Han ◽  
Bao Duong ◽  
Hongyi Li ◽  
...  
Keyword(s):  
B Cells ◽  
High Affinity ◽  
Affinity Ligand ◽  
High Affinity Ligand

scholarly journals RS-45041-190: a selective, high-affinity ligand for I2 imidazoline receptors

1995 ◽  
Vol 116 (2) ◽  
pp. 1737-1744 ◽  
Author(s):  
C.M. Brown ◽  
A.C. MacKinnon ◽  
W.S. Redfern ◽  
A. Williams ◽  
C. Linton ◽  
...  
Keyword(s):  
Imidazoline Receptors ◽  
High Affinity ◽  
Affinity Ligand ◽  
High Affinity Ligand

Isolation and characterization of some novel genes of the apolipoprotein A-I family in Japanese eel, Anguilla japonica

2011 ◽  
Vol 6 (4) ◽  
pp. 545-557 ◽  
Author(s):  
Malay Choudhury ◽  
Takahiro Oku ◽  
Shoji Yamada ◽  
Masaharu Komatsu ◽  
Keita Kudoh ◽  
...  
Keyword(s):  
Amino Acids ◽  
Molecular Weight ◽  
Amino Acid ◽  
Consensus Sequence ◽  
Structural Features ◽  
Lipid Binding ◽  
Japanese Eel ◽  
Amino Acid Sequences ◽  
Novel Genes ◽  
Isolation And Characterization

AbstractApolipoproteins such as apolipoprotein (apo) A-I, apoA-IV, and apoE are lipid binding proteins synthesized mainly in the liver and the intestine and play an important role in the transfer of exogenous or endogenous lipids through the circulatory system. To investigate the mechanism of lipid transport in fish, we have isolated some novel genes of the apoA-I family, apoIA-I (apoA-I isoform) 1–11, from Japanese eel by PCR amplification. Some of the isolated genes of apoIA-I corresponded to 28kDa-1 cDNAs which had already been deposited into the database and encoded an apolipoprotein with molecular weight of 28 kDa in the LDL, whereas others seemed to be novel genes. The structural organization of all apoIA-Is consisted of four exons separated by three introns. ApoIA-I10 had a total length of 3232 bp, whereas other genes except for apoIA-I9 ranged from 1280 to 1441 bp. The sequences of apoIA-Is at the exon-intron junctions were mostly consistent with the consensus sequence (GT/AG) at exon-intron boundaries, whereas the sequences of 3′ splice acceptor in intron 1 of apoIA-I1-7 were (AC) but not (AG). The deduced amino acid sequences of all apoIA-Is contained a putative signal peptide and a propeptide of 17 and 5 amino acid residues, respectively. The mature proteins of apoIA-I1-3, 7, and 8 consisted of 237 amino acids, whereas those of apoIA-I4-6 consisted of 239 amino acids. The mature apoIA-I10 sequence showed 65% identity to amino acid sequence of apoIA-I11 which was associated with an apolipoprotein with molecular weight of 23 kDa in the VLDL. All these mature apoIA-I sequences satisfied the common structural features depicted for the exchangeable apolipoproteins such as apoA-I, apoA-IV, and apoE but apoIA-I11 lacked internal repeats 7, 8, and 9 when compared with other members of apoA-I family. Phylogenetic analysis showed that these novel apoIA-Is isolated from Japanese eel were much closer to apoA-I than apoA-IV and apoE, suggesting new members of the apoA-I family.

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