scholarly journals On the Hydrazinolysis of Proteins and Peptides: A Method for the Characterization of Carboxyl-terminal Amino Acids in Proteins

1952 ◽  
Vol 25 (3) ◽  
pp. 214-218 ◽  
Author(s):  
Shiro Akabori ◽  
Ko Ohno ◽  
Kozo Narita
Keyword(s):  
Amino Acids ◽  
Terminal Amino ◽  
Carboxyl Terminal ◽  
Proteins And Peptides

scholarly journals RhoB prenylation is driven by the three carboxyl-terminal amino acids of the protein: Evidenced in vivo by an anti-farnesyl cysteine antibody

2000 ◽  
Vol 97 (21) ◽  
pp. 11626-11631 ◽  
Author(s):  
R. Baron ◽  
E. Fourcade ◽  
I. Lajoie-Mazenc ◽  
C. Allal ◽  
B. Couderc ◽  
...  
Keyword(s):  
Amino Acids ◽  
Terminal Amino ◽  
Carboxyl Terminal

Functional Analysis of a Peptide Sequence (Thr323 - Cys337) In ADAMTS13 Disintegrin-Like Domain Revealed Two Discontinuous Sequences Involved In the Interaction with VWF

Blood ◽  
2010 ◽  
Vol 116 (21) ◽  
pp. 2200-2200
Author(s):  
Atsuko Igari ◽  
Takanori Moriki ◽  
Terumichi Nakagawa ◽  
Yusuke Yamaguchi ◽  
Mitsuru Murata
Keyword(s):  
Amino Acids ◽  
Catalytic Activity ◽  
Synthetic Peptides ◽  
Inhibitory Effect ◽  
Peptide Sequence ◽  
Recent Analysis ◽  
Inhibitory Effects ◽  
Amino Terminal ◽  
Terminal Amino ◽  
Carboxyl Terminal

Abstract Abstract 2200 ADAMTS13 specifically cleaves multimeric von Willebrand factor (VWF) into smaller molecules to reduce its high reactivity with platelets. The disintegrin-like (D) domain, adjacent to the catalytic domain of ADAMTS13, plays an important role in the process of VWF cleavage. In this study, we aimed to elucidate critical peptide sequences in D-domain involved in the interaction with VWF. A series of partially overlapping peptide sequences, approximately 20 amino acids in length, covering the D-domain, were synthesized and the inhibitory effects on the catalytic activity of plasma ADAMTS13 was examined using FRETS-VWF73 assay. Consequently, some synthetic peptides were selected and the minimal length necessary for the inhibitory effect was determined as TFAREHLDMCQALSC (peptide323-337). Removal of the amino-terminal threonine diminished the inhibitory effect moderately, although deletion of the carboxyl-terminal cysteine abolished it completely. According to the amino acids alignment of ADAMTS family, this peptide sequence is not conserved, highlighting the specific role in the interaction with its substrate. From the recent analysis of crystal structure, amino-terminal half of the peptide323-337, TFAREHL (323-329), was disordered and designated as the variable (V) loop, which creates one of VWF-binding exosites (Akiyama, et al. Proc Natl Acad Sci USA. 2009; 106:19274-9). We hypothesized that the amino-terminal amino acids of the peptide323-337 contribute to VWF binding, whereas the carboxyl-terminal amino acids allow the structural stability of the peptide conformation. To evaluate the effect of carboxyl-terminal cysteine at 337, other synthetic peptides with alanine, serine, glycine or phenylalanine instead of the cysteine (C337A, C337S, C337G, or C337F) were tested about their inhibitory effects on the catalytic activity. Interestingly, C337A, C337S, C337G peptides exhibited slightly weaker inhibitory effects on VWF73 catalysis, although C337F peptide showed stronger inhibition than wild-type sequence, suggesting that the residue 337 regulates the characteristics of the peptide323-337. From the results of peptide screening, the amino- and carboxyl-terminal amino acids of the peptide323-337, TFAREHLDMCQALSC, likely play key roles in the inhibitory effects; therefore, the middle part of the sequence, HLDMC, was replaced by 5 alanines (AAAAA) or reversed sequence CMDLH. Surprisingly, the converted peptides still retained the equivalent level of inhibitory effects, indicating both sides of the amino- and carboxyl-terminal amino acids were especially significant in the interaction with VWF. In conclusion, we characterized the peptide sequence, TFAREHLDMCQALSC (323-337), in D-domain. The peptide clearly inhibited the cleavage of VWF73 and the both sides of amino- and carboxyl-terminal amino acids seemed especially important. The peptide sequence is supposed to bind to VWF for the precise cleavage in the process of proteolysis. By modifying this peptide sequence, such variant ADAMTS13 as gain-of-function recombinants might be developed, leading to an alternative anti-thrombotic drug. Disclosures: No relevant conflicts of interest to declare.

Characterization of a Histidine Rich Cluster of Amino Acids in the Cytoplasmic Domain of the Na+/H+ Exchanger

2000 ◽  
Vol 20 (3) ◽  
pp. 185-197 ◽  
Author(s):  
Pavel Dibrov ◽  
Rakhilya Murtazina ◽  
James Kinsella ◽  
Larry Fliegel
Keyword(s):  
Amino Acids ◽  
Fusion Protein ◽  
Cytoplasmic Domain ◽  
Intact Protein ◽  
Affinity Column ◽  
Rich Cluster ◽  
Protein Mutation ◽  
Conserved Sequence ◽  
Carboxyl Terminal

We examined the function of a highly conserved Histidine rich sequence ofamino acids found in the carboxyl-terminal of the Na+/H+exchanger (NHE1). A fusion protein containing the sequenceHYGHHH (540–545) and the balance of the carboxyl terminalof the protein did not bind calcium but bound to an immobilizedmetal affinity column and could be used to partially purify theexchanger protein. Mutation of the sequence to either HYGAAA orHYGRRR did not affect activity of the intact protein. Mutationto HHHHHH did not affect proton activation of the Na+/H+exchanger or localization but caused a decreased maximal velocitysuggesting that this conserved sequence is important in maximalactivity of the Na+/H+ exchanger.

scholarly journals Physicochemical characterization of carbamylated human serum albumin: an in vitro study

RSC Advances ◽  
2019 ◽  
Vol 9 (63) ◽  
pp. 36508-36516
Author(s):  
Asim Badar ◽  
Zarina Arif ◽  
Shireen Naaz Islam ◽  
Khursheed Alam
Keyword(s):  
Amino Acids ◽  
Human Serum Albumin ◽  
Serum Albumin ◽  
Human Serum ◽  
Physicochemical Characterization ◽  
In Vitro Study ◽  
Terminal Amino ◽  
Carbamyl Amino Acids

Carbamylation is an ubiquitous process in which cyanate (OCN−) reacts with the N-terminal amino or ε-amino moiety and generates α-carbamyl amino acids and ε-carbamyl-lysine (homocitrulline).

scholarly journals The Carboxyl Terminal Amino Acids of Rabbit γ-Globulin

1962 ◽  
Vol 237 (7) ◽  
pp. 2196-2200
Author(s):  
H.I. Silman ◽  
John J. Cebra ◽  
David Givol
Keyword(s):  
Amino Acids ◽  
Terminal Amino ◽  
Carboxyl Terminal
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