Purification of a base-specific ribonuclease Ru from Rhizopus niveus

1980 ◽  
Vol 58 (6) ◽  
pp. 489-493 ◽  
Author(s):  
Hiroyuki Horitsu ◽  
Noriyuki Takihi ◽  
Masahisa Sugiura ◽  
Mikio Tomoyeda
Keyword(s):  
Amino Acids ◽  
Aspartic Acid ◽  
Column Chromatography ◽  
Deae Cellulose ◽  
Disc Electrophoresis ◽  
Acetone Precipitation ◽  
Base Specificity ◽  
Terminal Amino ◽  
Carboxyl Terminal ◽  
Rhizopus Niveus

A base-specific ribonuclease (RNase) Ru (EC 3.1.27.5) was isolated and purified from Rhizopus niveus in a yield of 17% by the procedures of acetone precipitation, column chromatography on Duolite A-2, DEAE-cellulose, CM-cellulose, and 2′(3′)-aminohexyl-5′-UMP-agarose. The enzyme was shown to be homogeneous by polyacrylamide disc electrophoresis. The amino- and carboxyl-terminal amino acids of the enzyme were determined to be an arginine and an aspartic acid, respectively. The enzyme has a base specificity: it released only 3′-UMP from yeast RNA or poly(U) and, in addition, small amounts of 3′-CMP from poly(C).

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

scholarly journals Some properties of cytochrome b5 from liver microsomes of man, monkey, pig and chicken

1969 ◽  
Vol 115 (4) ◽  
pp. 849-856 ◽  
Author(s):  
F. G. Nóbrega ◽  
P. S. Araujo ◽  
M. Pasetto ◽  
I. Raw
Keyword(s):  
Amino Acids ◽  
Spectral Properties ◽  
Cytochrome B5 ◽  
Liver Microsomes ◽  
Gel Filtration ◽  
Deae Cellulose ◽  
Gradient Elution ◽  
Closely Related Species ◽  
Ammonium Sulphate Fractionation ◽  
Terminal Amino

1. Cytochrome b5 was released from liver microsomes of man, monkey, pig and chicken by incubation with a crude lipase preparation. 2. By using DEAE-cellulose chromatography, ammonium sulphate fractionation, Sephadex-gel filtration and a final gradient elution on DEAE-Sephadex A-50, cytochromes b5 were obtained from the four species studied, all possessing similar spectral properties. 3. Stokes radii of the cytochromes were measured by gel filtration. 4. N-Terminal amino acids for the different cytochromes were serine for man and monkey, alanine for pig and glycine for chicken. 5. Amino acid analyses of the cytochromes are presented. 6. Peptide ‘fingerprint’ patterns of tryptic digests of the different cytochromes are discussed and clearly show increasing similarity for more closely related species.

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.

scholarly journals Analysis of Fibrinolysis by the Use of Epsilon-Aminocaproic Acid: Preliminary Report

Blood ◽  
1960 ◽  
Vol 15 (5) ◽  
pp. 690-697 ◽  
Author(s):  
KATSUHIRO FUKUTAKE ◽  
KEIJI SHIDA ◽  
TOYOME ARAKAWA ◽  
KATSUJI KATO
Keyword(s):  
Amino Acids ◽  
Aspartic Acid ◽  
Preliminary Report ◽  
Inhibitory Action ◽  
Aminocaproic Acid ◽  
Terminal Amino ◽  
Epsilon Aminocaproic Acid

Abstract From the foregoing analysis of the inhibitory action of ε-aminocaproic acid in the fibrinolytic process, it is evident that the process involves two reaction phases, fibrinolysis and metafibrinolysis. Fibrinolysopeptide and metafibrin are split off from fibrin in fibrinolysis. The former is a peptide with N-terminal glycine, while the latter is a monochloracetic acid-soluble metaprotein having both aspartic acid and tyrosine as N-terminal amino acids.

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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