Monoclonal antibodies specific for bovine pancreatic asparagine synthetase. Production and use in structural studies.

Pfeiffer, N. E.; Mehlhaff, P M; Wylie, Dwane E.; Schuster, Sheldon M.

doi:10.1016/s0021-9258(17)36029-5

Cited by 18 publications

(2 citation statements)

References 28 publications

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“…Although total amounts of cross-linked products increased with increasing suberimidate concentration, the ratios of cross-linked products remained the same and no higher aggregate forms appeared. This same reactivity toward suberimidate (no higher aggregate forms even at 2.5 mg/mL suberimidate) has also been observed for several other proteins (Pfeiffer et al, 1986) with known quaternary structure.…”

Section: Resultssupporting

confidence: 78%

“…The quaternary structure standards for suberimidate cross-linking used in this study (hemoglobin and myoglobin) showed no artifactual subunit cross-linking (intermolecular cross-linking), even with suberimidate concentrations 67% higher than those used for phytochrome. Aldolase, /3-lactoglobins A and B, and carbonic anhydrase have also previously been (Pfeiffer et al, 1986) shown to demonstrate no artifactual cross-linking in conditions identical (except 1 mM EDTA was not used) with the ones for our controls (67% greater suberimidate concentration than was used in the phytochrome reaction).…”

Section: Discussionmentioning

confidence: 60%

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Spectral perturbations and oligomer/monomer formation in 124-kilodalton Avena phytochrome

Choi¹,

Kim²,

Kwon³

et al. 1990

Biochemistry

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We have studied the effects of pH, ionic strength, and hydrophobic fluorescence probes, 8-anilinonaphthalene-1-sulfonate (ANS) and bis-ANS, on the structure of intact (124-kDa) Avena phytochrome. The Pfr form of phytochrome forms oligomers in solution to a greater extent than the Pr form. Hydrophobic forces play a major role in the oligomerization of phytochrome, as suggested by fluorescence and monomerization by bis-ANS. However, electrostatic charges also take part in the phytochrome oligomerization. The partial proteolytic digestion patterns for the Pr and Pfr species are different, but binding of bis-ANS to the phytochrome abolishes this difference and yields an identical proteolytic peptide mapping for both spectral forms of phytochrome. This appears to result from bis-ANS binding at the carboxy-terminal domain, which induces monomerization of phytochrome oligomers. A second bis-ANS binding at an amino-terminal site blocks cleavage sites of trypsin and alpha-chymotrypsin. Bis-ANS especially blocks access of the proteases to the amino-terminal cleavage site that produces an early proteolytic product (114/118 kDa) on SDS gels. The bis-ANS binding does not, however, affect the proteolytic cleavage site that occurs in the hinge region between the two structural domains of phytochrome, the chromophore domain and the C-terminal non-chromophore domain. A chromophore binding site in the Pfr form is apparently exposed for preferential binding of bis-ANS, causing cyclization of the chromophore and bleaching of its absorbance at 730 nm. These observations have been discussed in terms of a photoreversible topographic change of the chromophore/apoprotein during the phototransformation of phytochrome.

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Section: Resultssupporting

confidence: 78%

Section: Discussionmentioning

confidence: 60%