PMID:1798706
| Citation |
Allemann, RK, Presnell, SR and Benner, SA (1991) A hybrid of bovine pancreatic ribonuclease and human angiogenin: an external loop as a module controlling substrate specificity? Protein Eng. 4:831-5 |
|---|---|
| Abstract |
A comparison of the sequences of three homologous ribonucleases (RNase A, angiogenin and bovine seminal RNase) identifies three surface loops that are highly variable between the three proteins. Two hypotheses were contrasted: (i) that this variation might be responsible for the different catalytic activities of the three proteins; and (ii) that this variation is simply an example of surface loops undergoing rapid neutral divergence in sequence. Three hybrids of angiogenin and bovine pancreatic ribonuclease (RNase) A were prepared where regions in these loops taken from angiogenin were inserted into RNase A. Two of the three hybrids had unremarkable catalytic properties. However, the RNase A mutant containing residues 63-74 of angiogenin had greatly diminished catalytic activity against uridylyl-(3'----5')-adenosine (UpA), and slightly increased catalytic activity as an inhibitor of translation in vitro. Both catalytic behaviors are characteristic of angiogenin. This is one of the first examples of an engineered external loop in a protein. Further, these results are complementary to those recently obtained from the complementary experiment, where residues 59-70 of RNase were inserted into angiogenin [Harper and Vallee (1989) Biochemistry, 28, 1875-1884]. Thus, the external loop in residues 63-74 of RNase A appears to behave, at least in part, as an interchangeable 'module' that influences substrate specificity in an enzyme in a way that is isolated from the influences of other regions in the protein. |
| Links | |
| Keywords |
Amino Acid Sequence; Animals; Binding Sites/genetics; Cattle; Endoribonucleases/chemistry; Endoribonucleases/genetics; Humans; Models, Molecular; Molecular Sequence Data; Protein Conformation; Proteins/chemistry; Proteins/genetics; Recombinant Fusion Proteins/chemistry; Recombinant Fusion Proteins/genetics; Ribonuclease, Pancreatic/chemistry; Ribonuclease, Pancreatic/genetics; Ribonucleases/chemistry; Ribonucleases/genetics; Sequence Homology, Nucleic Acid; Substrate Specificity/genetics |
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