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Nitrogen-15 spin-lattice relaxation times of amino acids in Neurospora crassa as a probe of intracellular environment

Kanamori, Keiko and Legerton, Timothy L. and Weiss, Richard L. and Roberts, John D. (1982) Nitrogen-15 spin-lattice relaxation times of amino acids in Neurospora crassa as a probe of intracellular environment. Biochemistry, 21 (20). pp. 4916-4920. ISSN 0006-2960. http://resolver.caltech.edu/CaltechAUTHORS:20151005-113554913

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Abstract

The nitrogen- 15 spin-lattice relaxation time, T_1, and the nuclear Overhauser enhancement (NOE) have been measured for intracellular glutamine, alanine, and arginine in intact Neurospora crassa mycelia to probe their various intracellular environments. The relaxations of ^(15)N_y of glutamine, ^(15)Nɑ of alanine, and ^(15)N ω,ω' of arginine in N. crassa were found, on the basis of their NOE values, to be predominantly the result of ^(15)N-H dipolar relaxation. These relaxations are therefore related to the microviscosities of the various environments and associations of the respective molecules with other cellular components that act to increase the effective molecular sizes. For ^(l5)Ny of glutamine in the cytoplasm, the intracellular T_1 (4.1 s) was only slightly shorter than that in the culture medium (4.9 s). This indicates that the microviscosity of the cytoplasm surrounding the glutamine molecules is not much greater than 1.3 cP. By contrast, for ^(15)Nω,ω,of arginine, which is sequestered in vacuoles containing polyphosphates, the intracellular T_1 (1.1 s) was only one-fourth of that in the medium (4.6 s). In model systems, the T_1, of ^(15)Nω,ω' in a 1 M aqueous solution of arginine containing 0.2 M pentaphosphate was 0.95 s, whereas in an isoviscous (2.8 cP) solution without pentaphosphate, the T1 was 1.8 s. These results suggest either that the vacuolar viscosity is substantially above 2.8 cP or that the ω,ω’-nitrogens of vacuolar arginine are associated with a polyanion, possibly polyphosphate. The implications of these results for the properties of the vacuolar interior are discussed in relation to the mechanism of amino acid compartmentation.


Item Type:Article
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URLURL TypeDescription
http://dx.doi.org/10.1021/bi00263a013DOIArticle
http://pubs.acs.org/doi/abs/10.1021/bi00263a013PublisherArticle
Additional Information:© 1982 American Chemical Society. Received February 29, 1982. This work was supported by US. Public Health Service Research Grant GM-11072 from the Division of General Medical Sciences (to K.K. and J.D.R.), by National Science Foundation Grant PCM78-19030 (to R.L.W.), and by US. Public Health Service National Research Award 07104 (to T.L.L.).
Funders:
Funding AgencyGrant Number
U.S. Public Health Service (USPHS)GM-11072
NSFPCM78-19030
U.S. Public Health Service (USPHS)07104
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Caltech Gates and Crellin Laboratories of Chemistry6598
Record Number:CaltechAUTHORS:20151005-113554913
Persistent URL:http://resolver.caltech.edu/CaltechAUTHORS:20151005-113554913
Official Citation:Nitrogen-15 spin-lattice relaxation times of amino acids in Neurospora crassa as a probe of intracellular environment Keiko Kanamori, Timothy L. Legerton, Richard L. Weiss, and John D. Roberts Biochemistry 1982 21 (20), 4916-4920 DOI: 10.1021/bi00263a013
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:60758
Collection:CaltechAUTHORS
Deposited By: Ruth Sustaita
Deposited On:05 Oct 2015 21:41
Last Modified:05 Oct 2015 21:41

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