Studies on the relationship between genes and enzymes

Abstract

The experimental work and theoretical discussion presented here suggests the following main conclusions.1. Argininosuccinase may be detected in extracts of a standard wild -type (SLA) and measurements may be made of some of the kinetics of its catalysis. I:t is considered likely, from these kinetic data and from the information on the behaviour of the enzyme on hydroxylapatite gel and on electrophoresis, that the enzyme is a single protein species.2. A report is given of an examination of some of the arg-10 mutants and of a heterokaryon between two of them.3.. The production and genetics of revertants of some of these arg-10 mutants, is described and it is suggested that at least two of the revertants (362r-1 and 362r-2) are the result of mutation(s) at or close to the arg-10 locus.4. The argininosuccinase formed by 362r-1 is described and it is proposed that the differences between this enzyme and that of the original wild-type (SLA) may be explained as the result of an alteration to the structure of the enzyme involving the active site.5. The argininosuccinase formed by 362r-2 is described and it is proposed that this enzyme differs both from that found in SLA and in 362r-1 and that this is also a reflection of an alteration in the structure of the active site of the enzyme.6. Some of the properties of argininosuccinase from K32 3-revertants are described and it is suggested that they may not differ in any way from the enzyme of SLA.7. A discussion is given of the growth of Neurospora crassa in culture and it is concluded that the growth -curves of the organism are complex and depend on the culture conditions.8. Measurements of argininosuccinase, argininosuccinic acid and arginine in cultures of SLA and 362r-1 are reported and the results obtained are explained in terms of a kinetic model of the arginine pathway in vivo. It is suggested that the concentration of arginine must always be independent of argininosuccinase concentration (at steady-state).9. In a discussion of the experiments and theory presented, the thesis is proposed that there are at least two major classes of catalyses, 'buffered" and "unbuffered", and that the genes affecting the enzymes concerned with these two types of catalyses will have distinct properties

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