Isolation, characterisation and cDNA sequencing of a new form of parvalbumin from carp semen

2014 ◽  
Vol 26 (8) ◽  
pp. 1117 ◽  
Author(s):  
Mariola A. Dietrich ◽  
Błażej Westfalewicz ◽  
Patrycja Jurecka ◽  
Ilgiz Irnazarow ◽  
Andrzej Ciereszko
Keyword(s):  
Amino Acid ◽  
Amino Acid Sequence ◽  
Molecular Mass ◽  
Seminal Plasma ◽  
Calcium Binding ◽  
Cdna Sequence ◽  
Full Length ◽  
Post Translational Modification ◽  
Preparative Electrophoresis ◽  
Full Length Cdna

Parvalbumins (Pv) are calcium-binding proteins present mainly in the muscle and nervous system where they act as a Ca2+ buffer. Our previous work demonstrated the presence of Pv-I in carp semen and indicated the presence of a second Pv (Pv-II). The purpose of the present work was to identify, purify and determine the full-length cDNA sequence of Pv-II from carp testis. Pv-II from seminal plasma was purified by ion-exchange chromatography (IEC) and preparative electrophoresis, while the Pv-II from spermatozoa was purified by IEC, gel filtration and preparative electrophoresis. The purified Pv-II was submitted to an analysis of molecular mass, isoelectric point (pI), amino-acid sequence and oligomerisation ability. The amino-acid sequence was used to construct primers and obtain the full-length cDNA sequence of seminal-specific Pv-II from carp testis. Analysis of the cDNA sequence indicated that carp-testis Pv-II was distinct from carp-muscle parvalbumins. Pv-II was distinct from Pv-I regarding sequence, molecular mass and pI. Both parvalbumins had the ability to form oligomers or to bind to other proteins. Carp seminal plasma had a protective effect against parvalbumin oligomerisation. Pv-II underwent post-translational modification such as n-acetylation and cysteinylation. The present study is the first to report the full-length cDNA sequence of parvalbumin from carp testis.

scholarly journals Isolation, characterization and sequence analysis of a full-length cDNA clone encoding acetohydroxy acid reductoisomerase from spinach chloroplasts

1991 ◽  
Vol 277 (2) ◽  
pp. 469-475 ◽  
Author(s):  
R Dumas ◽  
M Lebrun ◽  
R Douce
Keyword(s):  
Amino Acid ◽  
Amino Acid Sequence ◽  
Single Gene ◽  
Amino Acid Sequences ◽  
Full Length ◽  
Open Reading Frame ◽  
Amino Acid Residues ◽  
Protein Precursor ◽  
Reading Frame ◽  
Full Length Cdna

Acetohydroxy acid reductoisomerase (AHRI), the second enzyme in the parallel isoleucine/valine-biosynthetic pathway, catalyses an unusual two-step reaction in which the substrate, either 2-acetolactate or 2-aceto-2-hydroxybutyrate, is converted via an alkyl migration and an NADPH-dependent reduction to give 2,3-dihydroxy-3-methylbutyrate or 2,3-dihydroxy-3-methylvalerate respectively. We have isolated and characterized a full-length cDNA from a lambda gt11 spinach library encoding the complete acetohydroxy acid reductoisomerase protein precursor. The 2050-nucleotide sequence contains a 1785-nucleotide open reading frame. The derived amino acid sequence indicates that the protein precursor consists of 595 amino acid residues including a presequence peptide of 72 amino acid residues. The N-terminal sequence of the first 16 amino acid residues of the purified AHRI confirms the identity of the cDNA. The derived amino acid sequence from this open reading frame shows 23% identity with the deduced amino acid sequences of the Escherichia coli and Saccharomyces cerevisiae AHRI proteins. There are two blocks of conserved amino acid residues in these three proteins. One of these is a sequence similar to the ‘fingerprint’ region of the NAD(P)H-binding site found in a large number of NAD(P)H-dependent oxidoreductases. The other, a short sequence (Lys-Xaa-Xaa-Xaa-Xaa-Xaa-Xaa-Xaa-Ser-His-Gly-Phe) containing the amino acids lysine and histidine, could well be the catalytic site of the first step of the AHRI reaction. Southern-blot analysis indicated that AHRI is encoded by a single gene per haploid genome of about 7.5 kbp containing at least four introns.

scholarly journals The complete amino acid sequence of a pea (Pisum sativum) seed lipoxygenase predicted from a near full-length cDNA

1988 ◽  
Vol 253 (3) ◽  
pp. 915-918 ◽  
Author(s):  
P M Ealing ◽  
R Casey
Keyword(s):  
Amino Acid ◽  
Pisum Sativum ◽  
Amino Acid Sequence ◽  
Amino Acid Identity ◽  
Full Length ◽  
Predicted Amino Acid Sequence ◽  
Protein Coding ◽  
Full Length Cdna ◽  
Coding Regions

A near full-length cDNA for a pea (Pisum sativum) seed lipoxygenase was isolated and sequenced. It has a protein coding sequence (2583 bp), 5′ (59 bp) and 3′ (191 bp) non-coding regions, and a poly(A) tail (20 bp). The predicted amino acid sequence indicates a polypeptide of Mr 97,628 that shows about 86% amino acid identity with a soya-bean lipoxygenase 3 protein sequence [Yenofsky, Fine & Liu (1988) Mol. Gen. Genet. 211, 215-222]. The cDNA directs the transcription of mRNA that can be translated to give an anti-lipoxygenase-precipitable polypeptide in vitro.

scholarly journals Full Length cDNA Structure and Deduced Amino Acid Sequence of Human β-Hydroxy-5-Ene Steroid Dehydrogenase

1989 ◽  
Vol 3 (8) ◽  
pp. 1310-1312 ◽  
Author(s):  
Van Luu The ◽  
Yves Lachance ◽  
Claude Labrie ◽  
Gilles Leblanc ◽  
James L. Thomas ◽  
...  
Keyword(s):  
Amino Acid ◽  
Amino Acid Sequence ◽  
Full Length ◽  
Full Length Cdna ◽  
Steroid Dehydrogenase

scholarly journals Molecular cloning of chick cardiac muscle tensin. Full-length cDNA sequence, expression, and characterization.

1994 ◽  
Vol 269 (35) ◽  
pp. 22310-22319
Author(s):  
S.H. Lo ◽  
Q. An ◽  
S. Bao ◽  
W.K. Wong ◽  
Y. Liu ◽  
...  
Keyword(s):  
Cardiac Muscle ◽  
Molecular Cloning ◽  
Cdna Sequence ◽  
Full Length ◽  
Full Length Cdna

scholarly journals Three distinct anti-allergic drugs, amlexanox, cromolyn and tranilast, bind to S100A12 and S100A13 of the S100 protein family

1999 ◽  
Vol 338 (3) ◽  
pp. 583-589 ◽  
Author(s):  
Tsuyoshi SHISHIBORI ◽  
Yuhta OYAMA ◽  
Osamu MATSUSHITA ◽  
Kayoko YAMASHITA ◽  
Hiromi FURUICHI ◽  
...  
Keyword(s):  
Amino Acid ◽  
Amino Acid Sequence ◽  
Calcium Binding ◽  
Binding Proteins ◽  
S100 Protein ◽  
Reversed Phase ◽  
Molecular Probes ◽  
Calcium Binding Proteins ◽  
Ige Mediated ◽  
Human And Mouse

To investigate the roles of calcium-binding proteins in degranulation, we used three anti-allergic drugs, amlexanox, cromolyn and tranilast, which inhibit IgE-mediated degranulation of mast cells, as molecular probes in affinity chromatography. All of these drugs, which have different structures but similar function, scarcely bound to calmodulin in bovine lung extract, but bound to the same kinds of calcium-binding proteins, such as the 10-kDa proteins isolated in this study, calcyphosine and annexins I–V. The 10-kDa proteins obtained on three drug-coupled resins and on phenyl-Sepharose were analysed by reversed-phase HPLC. It was found that two characteristic 10-kDa proteins, one polar and one less polar, were bound with all three drugs, although S100A2 (S100L), of the S100 family, was bound with phenyl-Sepharose. The cDNA and deduced amino acid sequence proved our major polar protein to be identical with the calcium-binding protein in bovine amniotic fluid (CAAF1, S100A12). The cDNA and deduced amino acid sequence of the less-polar protein shared 95% homology with human and mouse S100A13. In addition, it was demonstrated that the native S100A12 and recombinant S100A12 and S100A13 bind to immobilized amlexanox. On the basis of these findings, we speculate that the three anti-allergic drugs might inhibit degranulation by binding with S100A12 and S100A13.

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