Summary
Polypeptides, synthesized from a mixture of amino acid amides by microwave heating during repeated hydration-dehydration cycles, showed hydrolase- and oxidoreductase-like catalytic activities. Poly(GAVDH), polypeptides synthesized from an equimolar mixture (each 0.1 M) of glycinamide,l-alaninamide,l-valinamide,l-aspartic acid α-amide, andl-histidinamide, catalyzed the hydrolysis of PNPAc. The hydrolytic rate of PNPAc with poly(GAVDH) was the quadruple of that ofl-histidine alone. Though the kcat values of different resulting polypeptides were 103–106 times less than those of native hydrolases, the Km value of the polypeptides further containing phenylalanine residues was nearly equal to that of the esterase. This result indicates the presence of hydrophobic interaction between a substrate and the polypeptides. Resulting polypeptides also showed catalytic activity for the reduction of ferricyanide ion [Fe(CN)3−] with NADH. The polypeptides seemed to have a strong affinity for adenine nucleotides, because the reaction was inhibited by adenine derivatives such as NAD+ and AppA. A hypothesis for the emergence of primitive protein enzymes is discussed.
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Abbreviations
- PNPAc:
-
p-nitrophenyl acetate
- PNPPr:
-
p-nitrophenyl propionate
- PNPBu:
-
p-nitrophenyl butyrate
- HPLC:
-
highperformance liquid chromatography
- TFA:
-
trifluoroacetic acid
- BSA:
-
bovine serum albumin
- Vmax :
-
maximum reaction velocity
- Km :
-
Michaelis constant
- kcat :
-
catalytic rate constant
- V′:
-
hydrolytic velocity per histidine residue contained in polypeptides
- 3-AcPyADH:
-
3-acetylpyridine adenine dinucleotide (reduced form)
- NMNH:
-
β-nicotinamide mononucleotide (reduced form)
- α-NADH:
-
α-nicotinamide adenine dinucleotide (reduced form)
- DCIP:
-
2,6-dichlorophenyl indophenol
- GTP:
-
guanosine 5′-triphosphate
- CTP:
-
cytidine 5′-triphosphate
- AppA:
-
P1, P2-diadenosine 5′-diphosphate
- ΔG′:
-
free energy change
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Ito, M., Handa, N. & Yanagawa, H. Synthesis of polypeptides by microwave heating II. Function of polypeptides synthesized during repeated hydration-dehydration cycles. J Mol Evol 31, 187–194 (1990). https://doi.org/10.1007/BF02109495
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DOI: https://doi.org/10.1007/BF02109495