scholarly journals Amino Acid Sequence of the Smaller Subunit of Conglutin ?, a Storage Globulin of Lupinus angustifolius

1977 ◽  
Vol 30 (2) ◽  
pp. 33 ◽  
Author(s):  
TC Elleman
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Amino Acid Sequence ◽  
Storage Protein ◽  
Storage Proteins ◽  
Lupinus Angustifolius ◽  
Amino Acid Residues ◽  
A Subunit

The amino acid sequence of the smaller subunit of conglutin y, the simplest of the three globulins from the seeds of Lupinus angusti/olius cv. Uniwhite, has been determined. The subunit was homogeneous and contained 154 amino acid residues, including five sulphur-containing amino acids-a considerably higher content than is found in most other legume storage proteins. There was no indication of the complexity experienced in studies of many other legume storage proteins. This is perhaps the first sequence of a subunit of a legume storage protein to be determined.

scholarly journals Hyaluronan-binding region of aggrecan from pig laryngeal cartilage. Amino acid sequence, analysis of N-linked oligosaccharides and location of the keratan sulphate

1992 ◽  
Vol 286 (3) ◽  
pp. 761-769 ◽  
Author(s):  
F P Barry ◽  
J U Gaw ◽  
C N Young ◽  
P J Neame
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Amino Acid Sequence ◽  
Signal Peptidase ◽  
Amino Acid Residues ◽  
Terminal Sequence ◽  
Binding Region ◽  
Keratan Sulphate ◽  
Laryngeal Cartilage ◽  
Hyaluronan Binding

The hyaluronan-binding region (HABR) was prepared from pig laryngeal cartilage aggrecan and the amino acid sequence was determined. The HABR had two N-termini: one N-terminal sequence was Val-Glu-Val-Ser-Glu-Pro (367 amino acids in total), and a second N-terminal sequence (Ala-Ile-Ser-Val-Glu-Val; 370 amino acids in total) was found to arise due to alternate cleavage by the signal peptidase. The N-linked oligosaccharides were analysed by examining their reactivity with a series of lectins. It was found that the N-linked oligosaccharide on loop A was of the mannose type, while that on loop B was of the complex type. No reactivity was detected between the N-linked oligosaccharide on loop B' and any of the lectins. The location of keratan sulphate (KS) in the HABR was determined by Edman degradation of the immobilized KS-containing peptide. The released amino acid derivatives were collected and tested for the presence of epitope to antibody 5-D-4. On the basis of 5-D-4 reactivity and sequencing yields, the KS chains are attached to threonine residues 352 and 357. There is no KS at threonine-355. This site is not in fact in G1, but about 16 amino acid residues into the interglobular domain. Comparison of the structure of the KS chain from the HABR and from the KS domain of pig laryngeal cartilage aggrecan was made by separation on polyacrylamide gels of the oligosaccharides arising from digestion with keratanase. Comparison of the oligosaccharide maps suggests that the KS chains from both parts of the aggrecan molecule have the same structure.

scholarly journals Molecular cloning and characterization of the cDNA coding for C4b-binding protein, a regulatory protein of the classical pathway of the human complement system

1985 ◽  
Vol 230 (1) ◽  
pp. 133-141 ◽  
Author(s):  
L P Chung ◽  
D R Bentley ◽  
K B Reid
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Amino Acid Sequence ◽  
Cdna Sequence ◽  
Binding Protein ◽  
Regulatory Protein ◽  
Protein A ◽  
Classical Pathway ◽  
Amino Acid Residues ◽  
Cloned Cdna

By using synthetic oligonucleotides as probes, plasmid clones containing portions of cDNA coding for human C4b-binding protein were isolated from a liver cDNA library. The entire amino acid sequence of the C4b-binding protein can be predicted from this study of the cloned cDNA when allied to a previous sequence study at the protein level [Chung, Gagnon & Reid (1985) Mol. Immunol. 22, 427-435], in which over 55% of the amino acid sequence, including the N-terminal 62 residues, was obtained. The plasmid clones isolated allowed the unambiguous determination of 1717 nucleotides of cDNA sequence between the codon for the 32nd amino acid in the sequence of C4b-binding protein and the 164th nucleotide in the 3′ non-translated region. The sequence studies show that the secreted form of C4b-binding protein, found in plasma, is composed of chains of apparent Mr 70 000 that contains 549 amino acid residues. Examination of the protein and cDNA sequence results show that there are at least two polymorphic sites in the molecule. One is at position 44, which can be glutamine or threonine, and the other is at position 309, which can be tyrosine or histidine. Northern-blot analysis indicated that the mRNA for C4b-binding protein is approx. 2.5 kilobases long. The N-terminal 491 amino acids of C4b-binding protein can be divided into eight internal homologous regions, each approx. 60 amino acids long, which can be aligned by the presence in each region of four half-cystine, one tryptophan and several other conserved residues. These regions in C4b-binding protein are homologous with the three internal-homology regions that have been reported to be present within the Ba region of the complement enzyme factor B and also to the internal-homology regions found in the non-complement beta 2-glycoprotein I.

scholarly journals Amino acid-sequence variability at the N-terminal extra piece of mouse immunoglobulin light-chain precursors of the same and different subgroups

1976 ◽  
Vol 157 (1) ◽  
pp. 145-151 ◽  
Author(s):  
Y Burstein ◽  
I Schechter
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Amino Acid Sequence ◽  
Light Chain ◽  
Variable Region ◽  
Amino Acid Residues ◽  
Free System ◽  
Methionine Residue ◽  
N Terminus ◽  
Mouse Myeloma

The proteins programmed in the wheat-germ cell-free system by the mRNA coding for the MOPC-63 mouse myeloma L (light) chain were labelled with six radioactive amino acids: [35S]methionine, [4,5-3H]leucine, [3,4-3H]proline, [3-3H]serine, [4,5-3H]isoleucine or [2,3-3H]alanine. Amino acid-sequence analyses showed that over 90% of the total cell-free product was one homogeneous protein, which corresponds to the MOPC-63 L-chain precursor. In this precursor an extra piece, 20 amino acid residues in length, precedes the N-terminus of the mature L chain. The extra piece contains one methionine residue at the N-terminus, six leucine residues, which are clustered in two triplets at positions 6, 7, 8 and 11, 12, 13, one proline residue at position 16, and one serine residue at position 18. The closely gathered leucine residues, as well as their abundance (30%), suggest that the extra-piece moiety is hydrophobic. In the precursors, the extra piece is coupled to the variable region of the L chain. Partial sequences of precursors of L chains of the same and different subgroups that were labelled with the above six radioactive amino acids indicate that the extra piece is part of the variable region. Thus the precursors of MOPC-63 and MOPC-321 L chains, which are of the same subgroup, have extra pieces of identical size (20 residues), and so far their partial sequences are also identical (see above). On the other hand, in the precursor of MOPC-41 L chain, which is of a different subgroup, the extra piece is 22 residues in length. Further, the sequence of the MOPC-41 extra piece differs in at least ten positions from sequences of the extra pieces of the precursors of MOPC-63 and MOPC-321 L chains.

scholarly journals The overexpression, purification and complete amino acid sequence of chorismate synthase from Escherichia coli K12 and its comparison with the enzyme from Neurospora crassa

1988 ◽  
Vol 251 (2) ◽  
pp. 313-322 ◽  
Author(s):  
P J White ◽  
G Millar ◽  
J R Coggins
Keyword(s):  
Escherichia Coli ◽  
Amino Acid ◽  
Amino Acid Sequence ◽  
Neurospora Crassa ◽  
Gel Filtration ◽  
Cross Linking ◽  
Amino Acid Residues ◽  
Chorismate Synthase ◽  
E Coli ◽  
A Subunit

The enzyme chorismate synthase was purified in milligram quantities from an overproducing strain of Escherichia coli. The amino acid sequence was deduced from the nucleotide sequence of the aroC gene and confirmed by determining the N-terminal amino acid sequence of the purified enzyme. The complete polypeptide chain consists of 357 amino acid residues and has a calculated subunit Mr of 38,183. Cross-linking and gel-filtration experiments show that the enzyme is tetrameric. An improved purification of chorismate synthase from Neurospora crassa is also described. Cross-linking and gel-filtration experiments on the N. crassa enzyme show that it is also tetrameric with a subunit Mr of 50,000. It is proposed that the subunits of the N. crassa enzyme are larger because they contain a diaphorase domain that is absent from the E. coli enzyme.

The Croonian Lecture, 1966 - The genetic code

1967 ◽  
Vol 167 (1009) ◽  
pp. 331-347 ◽  
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Nucleic Acid ◽  
Amino Acid Sequence ◽  
Genetic Code ◽  
Polypeptide Chain ◽  
Amino Acid Residues ◽  
Standard Set ◽  
Polypeptide Chains

Genes are made of nucleic acid. Enzymes are made of protein. The amino acid sequence of a particular protein is synthesized under instruction from a particular piece of nucleic acid. Each protein is made of one or more polypeptide chains, synthesized by condensing together amino acids, head to tail, with the elimination of water. A typical polypeptide chain is several hundred amino acid residues long. Nevertheless only twenty different kinds of amino acids are commonly found in proteins. This standard set of twenty is the same throughout nature. Nucleic acid is made of polynucleotide chains. The repeating unit of the chain is a sugar (ribose for RNA , deoxyribose for DNA ) connected to a phosphate. A base is joined on to each sugar. There are four common bases in nucleic acid. DNA usually has adenine, guanine, cytosine and thymine. In RNA thymine is replaced by uracil.

Amino acid sequence of penicillopepsin. IV. Myxobacter AL-1 protease II and Staphylococcus aureus protease fragments and homology with pig pepsin and chymosin

1976 ◽  
Vol 54 (10) ◽  
pp. 902-914 ◽  
Author(s):  
Anne Cunningham ◽  
Hsin-Min Wang ◽  
Stephen R. Jones ◽  
Alexander Kurosky ◽  
Leticia Rao ◽  
...  
Keyword(s):  
Amino Acids ◽  
Staphylococcus Aureus ◽  
Amino Acid ◽  
Amino Acid Sequence ◽  
Sequence Data ◽  
Amino Acid Residues ◽  
Terminal Sequence ◽  
Fungal Enzyme ◽  
Fungal Protease ◽  
And Staphylococcus Aureus

The digest of penicillopepsin (EC 3.4.23.7) with protease II from Myxobacter AL-1 gave five fragments which were separated on a Biogel P-100 column in 70% formic acid. The fragments were from 16 to 125 amino acids long. Two fragments were also isolated from a digest with a protease from Staphylococcus aureus. The analysis of these fragments by automatic sequencer gave a number of overlaps of the chymotryptic and thermolytic peptides. The available amino acid sequence data for penicillopepsin described in this paper and the accompanying papers (Kurosky, A. &Hofmann, T.: Can. J. Biochem. 54, 872 (1976); Rao, L. &Hofmann, T.: Can. J. Biochem. 54, 885 (1976); Harris, C. I., Rao, L., Shutsa, P., Kurosky, A. &Hofmann, T.: Can. J. Biochem. 54, 895 (1976)) have been combined and yield 15 fragments which range in lengths from 3 to 112 amino acid residues. These unique fragments account for virtually all the amino acids of the fungal protease. Four of the fragments with a total of 194 residues (about 60% of the molecule) have been aligned with corresponding sections of pig pepsin (EC 3.4.23.1) and with part of the N-terminal sequence available for calf chymosin (EC 3.4.23.4). In the alignments about 37% of the residues in the fungal enzyme are identical with at least one of the mammalian enzymes. An additional 20% are chemically similar. These results, together with previously reported active-site directed modifications, show conclusively that penicillopepsin is an evolutionary homologue of the mammalian acid proteases.

Molecular cloning of cDNA for porcine prolactin precursor

1990 ◽  
Vol 4 (2) ◽  
pp. 135-142 ◽  
Author(s):  
Y. Kato ◽  
T. Hirai ◽  
T. Kato
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Amino Acid Sequence ◽  
Signal Sequence ◽  
Cdna Clones ◽  
Amino Acid Residues ◽  
Ancestral Gene ◽  
Wide Range ◽  
Hydrophilic Residues ◽  
Ovine Prolactin

ABSTRACT Porcine prolactin cDNA clones were screened using antiserum against ovine prolactin from a cDNA library of porcine anterior pituitary, and their nucleotide sequences were determined by the chain-termination method. The nucleotide sequence of the 5′ untranslated region and part of the signal peptide region were determined by direct RNA sequencing with reverse transcriptase. The composite sequence of 957 nucleotides showed a signal sequence of 30 amino acids and a further 199 amino acids corresponding to the mature prolactin molecule. The predicted sequence confirmed the amino acid sequence determined previously by direct protein analysis, except for one amide form at residue 122 (Gln instead of the reported Glu). Northern blot analysis showed that the length of the porcine prolactin mRNA was about 1·1 kb. The porcine prolactin amino acid sequence showed 81, 80, 64, 62, 80 and 31% homology with human, bovine, rat, mouse, chick and salmon forms respectively. The identical amino acid residues showed marked clustering in four domains, two of which are highly conserved throughout a wide range of species. The hydropathy and secondary structure of porcine prolactin were analysed and compared with those of porcine GH, which shares the same ancestral gene. The two highly conserved regions of both hormones showed similar hydrophilicity, and the predicted secondary structures indicated that these regions in each hormone form different structures with differences in extension of the hydrophilic residues outside the molecule.

Cloning of pea storage protein genes

1984 ◽  
Vol 304 (1120) ◽  
pp. 323-332 ◽  
Keyword(s):  
Amino Acid ◽  
Amino Acid Sequence ◽  
Storage Protein ◽  
Genomic Dna ◽  
Storage Proteins ◽  
Amino Acid Sequences ◽  
Chromosome 7 ◽  
Messenger Rnas ◽  
Specific Storage ◽  
Storage Protein Genes

Vicilin and legumin are the major storage proteins of Pisum sativum . Complementary DNAs (cDNAs) have been produced from poly(A) + mRNA isolated from developing seeds and specific storage protein cDNAs cloned into pBR322. The amino acid sequences predicted from the cDNA sequences have been compared with the actual amino acid sequences derived from the purified protein subunits. These comparisons have confirmed that the legumin α and β subunits as initially synthesized are covalently joined together and that a small peptide is subsequently removed by endoproteolysis to give the disulphide linked subunits of the mature seed legumins. Similar comparisons between the predicted amino acid sequence of vicilin cDNA clones and the amino acid sequence determined on the isolated subunits has shown that some of the 50000 M r type subunits are subsequently cleaved to give three subunits as products, i.e. polypeptides of 19000 M r (α), 13500 M r (β) and 12500 M r or 16000 M r if glycosylated (γ). In addition to these three subunits, cleavage at one or other of the two potential cleavage sites, results in a 33000 M r polypeptide (α + β) and a 31000 M r polypeptide tentatively identified as β + γ. The presence of the sequence Lys-Glu-Asn leads to cleavage on the carboxy side of Asn at the β :γ cleavage site whereas the sequence Gly-Leu-Arg does not lead to cleavage. Comparable sequence data for the α: β processing site do not exist. Comparisons of the cDNA and amino acid sequence disclose the presence of a 15 or 16 amino acid residue vicilin leader sequence as well as a 12 amino acid residue C-terminal peptide which is also removed. The codon usage of the messenger RNAs for the storage proteins are similar to those of other plant proteins and differ somewhat from animal messenger RNAs. Complementary DNAs for specific storage proteins when used to probe different restriction enzyme digests of pea genomic DNA reveal the presence of a small number of legumin and vicilin coding sequences (two to five for legumin and three to seven for vicilin) that occur as single copies except for one vicilin sequence present in two to three copies. Genetic mapping experiments using whole plants locate both the main legumin and the vicilin genes on chromosome 7. The main legumin subunits are coded by genes located at a single Mendelian locus Lg-1 located on the short arm of chromosome 7 very close to the rub locus and the vicilin gene is located 16 map units away close to the r locus. Gene libraries prepared with size fractionated partial restriction enzymic digests of pea genomic DNA ligated into both phage λ L47 and phage λ gt wes have led to the isolation of at least three similar but different legumin genomic sequences. Comparison of the λ and cDNA legumin clones suggests the presence of at least one intron in the former. Legumes in general contain two major seed storage protein types, vicilin and legumin (Derbyshire et al. 1976). Seeds of Pisum sativum (L) have significant amounts of both proteins and since a considerable body of knowledge exists about pea physiology and genetics, this species is a good choice for the study of storage protein genes. Peas are also one of the world’s major legume crops. Since the storage proteins are found only in the tissues of the developing seed (Millerd 1975) and then only in significant amounts during the middle and late stages of development, it was suspected from the onset that the genes responsible for the storage protein would belong to the class of developmentally regulated genes, i.e. those that are only switched on in specific tissues over restricted periods of time.

scholarly journals Cloning and expression of diadenosine 5′,5‴-P1,P4-tetraphosphate hydrolase from Lupinus angustifolius L

1998 ◽  
Vol 329 (2) ◽  
pp. 313-319 ◽  
Author(s):  
Danuta MAKSEL ◽  
Andrzej GURANOWSKI ◽  
C. Steven ILGOUTZ ◽  
Arthur MOIR ◽  
G. Michael BLACKBURN ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Amino Acids ◽  
Amino Acid ◽  
Amino Acid Sequence ◽  
Sequence Similarity ◽  
Lupinus Angustifolius ◽  
Cloning And Expression ◽  
Lupinus Luteus ◽  
Higher Plant ◽  
Polyadenylated Rna

The first isolation, cloning and expression of cDNA encoding an asymmetric diadenosine 5ʹ,5‴P1,P4-tetraphosphate pyrophosphohydrolase (Ap4A hydrolase) from a higher plant is described. Ap4A hydrolase protein was purified from seeds of both Lupinus luteus and Lupinus angustifolius and partially sequenced. The Ap4A hydrolase cDNA was cloned from L. angustifolius cotyledonary polyadenylated RNA using reverse transcription and PCR with primers based on the amino acid sequence. The cDNA encoded a protein of 199 amino acids, molecular mass 22982 Da. When expressed in Escherichia coli fused to a maltose-binding protein, the enzyme catalysed asymmetric cleavage of Ap4A to AMP and ATP which was inhibited at concentrations of F- as low as 3 μM. These are properties characteristic of Ap4A hydrolase (asymmetrical) (EC 3.6.1.17). Comparison of the Ap4A hydrolase sequences derived from the four known cDNAs from pig, human, lupin and fission yeast showed that, like the mammalian hydrolase, the lupin enzyme possesses a Mut T motif but no other significant similarities. No sequence similarity to the human fragile histidine triad protein, as found in the Ap4A hydrolase from Schizosaccharomyces pombe, was detected in the Ap4A hydrolase from lupin.

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