Comparison of the zinc binding domains in the 7S nerve growth factor and the zinc-insulin hexamer

Dunn, Michael F.; Pattison, Scott E.; Storm, Michael C.; Quiel, Edward

doi:10.1021/bi00545a017

Cited by 52 publications

(51 citation statements)

References 21 publications

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“…When the [Zn]/[In]' ratio is 60.33, the hexameric state predominates; at higher ratios, there is a pronounced tendency to form higher aggregates and to crystallize or precipitate (Cunningham et al, 1955;Schlichtkrull, Fredericq, 1956;Grant et al, 1972;Jeffrey, 1974;Goldman & Carpenter, 1974;Milthorpe et al, 1977). Earlier work from this laboratory (Dunn et al, 1980) has shown that, in mixtures of zinc and insulin with a [Zn2+]/[In] ratio below 0.33, the reactivity of the zinc ions toward a large excess of the tridentate chelator 2,2',2"-terpyridine (terpy) is indeed consistent with the assignment of the metal ion environment to that of the crystallographically identified His-B 10 sites. Fourier-transform 'I3Cd N M R experiments with 1'3Cd2+-substituted insulin have shown that Cd2+ substitution for Zn2+ results in the formation of a cadmium-insulin species presumed to be (In)6(Cd2+)2Cd2+; the two classes of highaffinity Cd2+ sites have been assigned as His-B10 sites and a new site assigned to the Glu-B13 carboxylates located at the center of the hexamer.2 Metal ion substitution experiments +This work was supported by NIH Grant PHS-1-ROI-AM31138 and by a grant-in-aid to N.C.K.…”

supporting

confidence: 68%

“…The existence in solution of two (identical) high-affinity zinc sites on the insulin hexamer, i.e., the crytallographically identified His-B 10 sites, has been demonstrated (Fredericq, 1956;Grant et al, 1972;Milthorpe et al, 1977;Dunn et al, 1980;Sudmeier et al, 1981). Since a large excess of insulin relative to Zn2+ must favor binding to these sites, we assign the 412-nm-absorbing species to the ternary complex in which the monoanion of PAR is coordinated to Zn2+ residing in the His-B10 site.…”

Section: -80 Pm)mentioning

confidence: 96%

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Effects of calcium ion on ternary complexes formed between 4-(2-pyridylazo)resorcinol and the two-zinc insulin hexamer

Kaarsholm¹,

Dunn²

1987

Biochemistry

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As a means for probing the microenvironment of zinc in the insulin hexamer and to investigate the effects of calcium ion on the assembly and the structure of the two-zinc insulin hexamer, the thermodynamics and kinetics of the reaction between the chromophoric divalent metal ion chelator 4-(2-pyridylazo)resorcinol (PAR) and zinc-insulin have been investigated over a wide range of conditions. For [PAR]0 much greater than [Zn2+]0 and [Zn2+]/[In] less than or equal to 0.33, the reaction leads to the sequestering and ultimate removal of all of the insulin-bound Zn2+; for [Zn2+]0 much greater than [PAR]0, two stable ternary complexes are formed where Zn2+ has ligands derived from PAR as well as from hexameric insulin. For [Zn2+]/[In] ratios below 0.33, the equilibrium distribution between the two ternary complexes is dependent on the [Zn2+]/[In] ratio. One of the complexes is assigned to the monoanion of PAR coordinated to Zn2+ that resides in a His-B10 site. The other complex is proposed to involve the coordination of (PAR)Zn to the site formed by the alpha-NH2 group of Phe-B1 and the gamma-carboxylate ion of Glu-A17 across the dimer-dimer interface on the surface of the hexamer. With either PAR or zinc-insulin in large excess, the kinetics of the PAR optical density changes are remarkably similar and biphasic. The faster step is first order in PAR and first order in insulin-bound Zn2+ (k congruent to 3 X 10(3) M-1 s-1) and involves the formation of an intermediate in which PAR is coordinated to insulin-bound zinc at the His-B10 site.(ABSTRACT TRUNCATED AT 250 WORDS)

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supporting

confidence: 68%

Section: -80 Pm)mentioning

confidence: 96%

Effects of calcium ion on ternary complexes formed between 4-(2-pyridylazo)resorcinol and the two-zinc insulin hexamer

Kaarsholm¹,

Dunn²

1987

Biochemistry

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“…For measurement of the effect of metal ions on ALAD activity, the crude lysates, prepared as above, were passed through a BioRad 25 mm disc with Chelex Chelating Ion Exchange Resin, according to the manufacturer's instructions. This procedure removes all loosely bound metal ions [8].…”

Section: Enzyme Assaysmentioning

confidence: 99%

“…Crude tissue extracts from various organisms were prepared and a sample assayed for ALAD activity. The rest was passed through a BioRad Chelex Chelating Ion Exchange Resin as described above, which effectively removes the majority of loosely bound metal ions [8]. When this is done with pea leaf extract, the level of ALAD activity is severely reduced ( Table 1), but this can be restored to the original specific activity by titration with Mg 2+ ions.…”

Section: Metal Requirement Of Cyanobacterial Aladmentioning

confidence: 99%

Cloning and characterisation of genes for tetrapyrrole biosynthesis from the cyanobacterium Anacystis nidulans R2

et al. 1994

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The genes for 5-aminolevulinic acid dehydratase (ALAD) and uroporphyrinogen III synthase (UROS), two enzymes in the biosynthetic pathway for tetrapyrroles, were independently isolated from a plasmid-based genomic library of Anacystis nidulans R2 (also called Synechococcus sp. PCC7942), by their ability to complement Escherichia coli strains carrying mutations in the equivalent genes (hemB and hemD respectively). The identity of the genes was confirmed by comparing the appropriate enzyme activities in complemented and mutant strains. Subclones of the original plasmids that were also capable of complementing the mutants were sequenced. The inferred amino acid sequence of the cyanobacterial HemB protein indicates a significant difference in the metal cofactor requirement from the higher-plant enzymes, which was confirmed by overexpression and biochemical analysis. The organisation of the cyanobacterial hemD locus differs markedly from other prokaryotes. Two open reading frames were found immediately upstream of hemD. The product of one shows considerable similarity to published sequences from other organisms for uroporphyrinogen III methylase (UROM), an enzyme involved in the production of sirohaem and cobalamins (including vitamin B-12). The product of the other shows motifs which are similar to those found in proteins responsible for metabolic regulation in yeast and indicates that this family of transcription control proteins, which has previously been reported only from eukaryotes, is also represented in prokaryotes.

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“…Previous experiments suggested that the octapeptide sequence does not affect 7S NGF formation [Mobley et al, 1976]. However, the stability of 7S NGF is greatly dependent upon the presence of zinc which binds to 7S NGF with an apparent stoichiometry of two zinc ions for each complex [Pattison and Dunn, 1975], As His-8 has been implicated by homology with insulin as the residue in (3NGF coordi nating zinc [Dunn et al, 1980], the possibility exists that the octapeptide region, specifi cally His-8, mediates the zinc-conferred sta bility of 7S NGF. This was shown not to be true as the results in figure 4 demonstrate.…”

Section: Protection O F Pngf By a Subunit Y Subunit And 7s Ngf Againmentioning

confidence: 99%

Subunit Interactions of the Nerve and Epidermal Growth Factor Complexes: Protection of the Biological Subunit from Proteolytic Modification

Nichols¹,

Shooter²

1985

Dev Neurosci

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The nerve growth factor dimer (βNGF) can undergo two proteolytic modifications, one near the amino terminus where a unique histidine/methionine bond is cleaved and the other at the carboxy terminus releasing the terminal arginine residue. The modification near the amino terminus, releasing the first eight amino acids as an octapeptide, occurs as the result of the action of a specific endopeptidase present in the submaxillary gland. This same enzyme was found to be present in high concentration in epinephrine-stimulated saliva (20 mM) and was isolated from this source. The subunit interactions in 7S NGF were considered by assessing the protection afforded by each subunit, individually and in combination, against these two proteolytic modifications. Similar experiments were attempted with high molecular weight epidermal growth factor (HMW-EGF), an analogous growth factor complex that can experience a single modification, the release of arginine by carboxypeptidase B from the carboxy terminus of its biological subunit, low molecular weight EGF (LMW-EGF). The amino terminal octapeptide of βNGF is protected by both the α and γ subunits of 7S NGF and its loss has no effect on 7S NGF complex stabilization by zinc. The carboxy terminal arginine is protected only by the γsubunit. A scheme depicting the subunit interactions in 7S NGF is presented.

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Comparison of the zinc binding domains in the 7S nerve growth factor and the zinc-insulin hexamer

Cited by 52 publications

References 21 publications

Effects of calcium ion on ternary complexes formed between 4-(2-pyridylazo)resorcinol and the two-zinc insulin hexamer

Effects of calcium ion on ternary complexes formed between 4-(2-pyridylazo)resorcinol and the two-zinc insulin hexamer

Cloning and characterisation of genes for tetrapyrrole biosynthesis from the cyanobacterium Anacystis nidulans R2

Subunit Interactions of the Nerve and Epidermal Growth Factor Complexes: Protection of the Biological Subunit from Proteolytic Modification

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