Nuclease-T: an active derivative of staphylococcal nuclease composed of two noncovalently bonded peptide fragments.

Taniuchi, Hiroshi; Anfinsen, Christian B.; Sodja, Ann

doi:10.1073/pnas.58.3.1235

Cited by 121 publications

(37 citation statements)

References 7 publications

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“…Thermolytic digestion was performed in 0.2 % NH4HC03 pH 8.3 at 37 "C for about 20 h, and digestion with chymotrypsin in the same conditions for 3 -5 h. Subtilisin digestion was carried out either in 0.7% NH4HC03 or in 0.2 M N-ethylmorpholine acetate pH 8.0 at 37 "C for about 16 h. Digestions with carboxypeptidases A and B [14,15] and pronase [16] were carried out as described previously.…”

Section: Enzymic Hydrolysis Of Peptidesmentioning

confidence: 99%

The Amino‐Acid Sequence of Leghemoglobin Component a from Phaseolus vulgaris (Kidney Bean)

Lehtovaara¹,

Ellfolk²

1975

European Journal of Biochemistry

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1. Leghemoglobin component a from Phaseolus vulgaris (kidney bean) was digested with trypsin; 15 tryptic peptides and free lysine were purified and the amino acid sequences of the peptides determined.2. The internal order of the tryptic peptides was determined by the bridge peptides obtained from the thermolytic digest and the dilute acid hydrolyzate of kidney bean leghemoglobin a ; 12 thermolytic peptides and two acid hydrolysis peptides were purified and the sequences were partially or completely determined.3. The complete amino acid sequence of kidney bean leghemoglobin a is compared to that of leghemoglobin a from soybean (Glycine max) and to some animal globins. As regards sequence, the kidney bean globin has 79 % identity with the soybean globin and 21 % identity with human hemoglobin y-chain. Seven of the 14 amino acid residues common to most globins are found in the kidney bean globin. Trp-I 5 and Tyr-145 are evolutionarily conserved in this globin, which confirms the concept of a common origin of animal and plant globins.The only globin currently known in plant kingdom is the single-chain leghemoglobin from leguminous root nodules. It has many functional properties in common with animal hemoglobins and myoglobins. The evolution of leghemoglobin is not understood. It may possibly be elucidated by comparing the primary structures of several different leguminous globins with animal globins. Knowledge of the primary structures of related proteins is also of importance when studying the relationship of structure to function and chemical properties. A good deal of information about animal globins is now available. For soybean leghemoglobin, the best known of the plant globins, the oxygen binding characteristics, the main principles of its function [1-51 and the primary structure [6-101 have been clarified. This work with the primary structure of Phaseolus vulgaris (kidney bean) leghemoglobin component a has been preceded by the characterization of the

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Section: Enzymic Hydrolysis Of Peptidesmentioning

confidence: 99%

The Amino‐Acid Sequence of Leghemoglobin Component a from Phaseolus vulgaris (Kidney Bean)

Lehtovaara¹,

Ellfolk²

1975

European Journal of Biochemistry

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“…Proteins reconstituted from fragments produced by limited proteolytic cleavage or genetic methods have been studied under both in vivo and in vitro conditions [9][10][11][12][13]. Successful reconstitution implies that the internal complementarity must be highly self-selective to overcome competing interactions with other protein sequences [14].…”

Section: Introductionmentioning

confidence: 99%

Reconstitution of the enzyme AroA and its glyphosate tolerance by fragment complementation

et al. 2006

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5-Enolpyruvylshikimate-3-phosphate (EPSP) synthase (AroA) is a key enzyme in the aromatic amino acid biosynthetic pathway in microorganisms and plants, and is the target of the herbicide glyphosate. Glyphosate tolerance activity of the enzyme could be obtained by natural occurrence or by site-directed mutagenesis. A functional Pseudomonas putida AroA was obtained by co-expression of two protein fragments AroA P. putida -N210 and AroA P. putida -C212 in Escherichia coli aroA mutant strain AB2829. From sequence analysis, the equivalent split site on E. coli AroA was chosen for further study. The result indicated that functional E. coli AroA could also be reconstituted from two protein fragments AroA E. coli -N218 and AroA E. coli -C219, under both in vivo and in vitro conditions. This result suggested that the fragment complementation property of this family of enzyme may be general. Additional experiments indicated that the glyphosate tolerance property of AroA could also be reconstituted in parallel with its enzyme activity. The implication of this finding is discussed.

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“…The peptides 6-48 (or 49) and 49 (or 50) to 149 are enzymatically inactive and have random coil structures. When recombined to yield "nuclease T", the native conformation appears to be restored together with loo/,, of the original activity [33,34]. Most decisively against the theory of sequential folding from the amino end is the observation that cleaving residues 127-149 off the enzyme destroys t'he native conformation of the remaining chain and produces a random coil structure.…”

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confidence: 99%

X‐Ray Analysis, Structure and Function of Enzymes

Perutz

1969

European Journal of Biochemistry

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Lectures named after great scientists are generally instituted in honour of the dead. FEBS is to be congratulated on honouring in this way a colleague who is still active and whom we all admire for the work he does and the man he is. I feel deeply touched trhat I, an X-ray crystallographer and not a bio- If we follow Monod and elevate myoglobin and haemoglobin to the rank of honorary enzymes, then 1968 is the tenth anniversary of the publication of

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Nuclease-T: an active derivative of staphylococcal nuclease composed of two noncovalently bonded peptide fragments.

Cited by 121 publications

References 7 publications

The Amino‐Acid Sequence of Leghemoglobin Component a from Phaseolus vulgaris (Kidney Bean)

The Amino‐Acid Sequence of Leghemoglobin Component a from Phaseolus vulgaris (Kidney Bean)

Reconstitution of the enzyme AroA and its glyphosate tolerance by fragment complementation

X‐Ray Analysis, Structure and Function of Enzymes

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