Photoaffinity labeling of the hormone binding site of neurophysin.

Abercrombie, David M.; McCormick, William; Chaiken, Irwin M.

doi:10.1016/s0021-9258(18)34917-2

Cited by 30 publications

(11 citation statements)

References 37 publications

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“…The effects of excision of residues 1-8 on Tyr-49 are potentially sufficient to account for the diminished peptidebinding ability of the des-1-8 protein. Tyr-49 has had an uneven history with respect to its proposed relationship to the peptide-binding site [e.g., see Balaram et al (1973) and Lord & Breslow (1980)], but NMR and affinity-labeling evidence now clearly supports its proximity to this site (Abercrombie et al, 1982b;Peyton & Breslow, 1985). Accordingly, conformational alterations in the vicinity of Tyr-49 have the potential for significant repercussions on binding affinity.…”

Section: Discussionmentioning

confidence: 99%

“…Additionally, relatively little is known about the peptidebinding site. Early and recent studies provide increasing evidence for the proximity of the single neurophysin tyrosine, Tyr-49, to the peptide site (Balaram et al, 1973;Abercrombie et al, 1982b;Peyton & Breslow, 1985); Glu-31 has also been implicated (Walter & Hoffman, 1973). An unsolved question is the relative role of neurophysin's internally duplicated sequences (12-31 and 60-75) in binding and self-association (Breslow, 1979;Cohen et al, 1979).…”

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confidence: 99%

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Dimerization of native and proteolytically modified neurophysins as monitored by proton magnetic resonance spectroscopy: proximity of tyrosine-49 to the subunit interface

Peyton¹,

Sardana²,

Breslow³

1986

Biochemistry

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Neurophysin is a self-associating protein in which peptide-hormone binding and dimerization are thermodynamically linked. The structural basis of the linkage is unknown. We have studied the dimerization of bovine neurophysin I and two proteolytically modified derivatives by proton nuclear magnetic resonance spectroscopy in order to identify residues at the intersubunit contact regions and to evaluate the origin of the reported loss of dimerization associated with tryptic excision of residues 1-8. The concentration dependence at neural pH of the spectra of native neurophysin and des-90-92-neurophysin demonstrated a finite set of dimerization-sensitive resonances that included the ring protons of Tyr-49. Using these to monitor dimerization, we confirmed predictions of a large increase in the dimerization constant associated with carboxyl protonation. By the same criteria, dimerization of the des-1-8 protein, in disagreement with earlier reports, was found to be undiminished relative to that of the native protein. However, spectral changes in the Tyr-49 ring ortho proton region associated with dimerization of the des-1-8 protein differed significantly from those in the native protein and indicated an altered conformation of the des-1-8 dimer apparently restricted to the vicinity of Tyr-49. The results are shown to place Tyr-49 adjacent to both the intersubunit contact region and the 1-8 sequence in the native protein, loss of stabilizing interactions with 1-8 leading to altered interactions of Tyr-49 with the subunit interface. Because Tyr-49 is also close to the peptide-binding site, this arrangement spatially links the peptide-binding and dimerization sites of neurophysin.(ABSTRACT TRUNCATED AT 250 WORDS)

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Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

Dimerization of native and proteolytically modified neurophysins as monitored by proton magnetic resonance spectroscopy: proximity of tyrosine-49 to the subunit interface

Peyton¹,

Sardana²,

Breslow³

1986

Biochemistry

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“…Performic acid oxidation of native and trypsin-digested neurophysins was performed by routine procedures (Chaiken & Hough, 1980;Abercrombie et al, 1982b).…”

Section: Methodsmentioning

confidence: 99%

“…Amino acid compositions were determined on a Beckman Model 121 M analyzer. Ratios of moles of residues per mole of protein were computed as described previously (Abercrombie et al, 1982b).…”

Section: Methodsmentioning

confidence: 99%

“…A partially cohesive view of the hormone binding surface has been deduced by a combination of studies such as examination of hormone and peptide ligand binding specificities (Breslow & Weis, 1972;Breslow et al, 1973;Gamier et al, 1973; Glasel et al, 1976;Nicolas et al, 1976), spectroscopic changes induced by binding of unlabeled ligands (Balaram et al, 1973;Griffin et al, 1973;Sur et al, 1979) and spin-labeled ligands (Lord & Breslow, 1980), effects of chemical modifications (Walter & Hoffman, 1973;Schlesinger et al, 1979), and photoaffinity labeling (Abercrombie et al, 1982b). The proximity to the hormone binding site of both a carboxylate anion and Tyr-49 has been discussed (Breslow & Garguilo, 1977;Nicolas et al, 1978; Abercrombie et al, 1982b). In contrast to data for the hormone binding site, little information is yet available for the neurophysin binding surface participating in self-association.…”

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confidence: 99%

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Effects of limited tryptic proteolysis of bovine neurophysins on molecular properties of hormone binding, self-association, and antigenicity

Abercrombie¹,

Angal²,

Sequeira³

et al. 1982

Biochemistry

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Limited tryptic fragmentation of disulfide-intact bovine neurophysins I and II (NP-I and -II, respectively) has been found to cause selective disruption of both hormone binding and neurophysin self-association. Loss of binding interactions, measured as a loss of ability to stimulate retardation of 125I-labeled neurophysin on Met-Tyr-Phe-amino-butylaminoagarose, is complete within 3 h at 37 degrees C. Reverse-phase high-performance liquid chromatography (HPLC) analysis of tryptic digests of neurophysin I allows detection of two major protein products and the peptide fragment 1-8. Release of the latter N-terminal piece occurs at about the same rate as loss of binding interactions. Reverse-phase HPLC elution behavior before and after performic acid oxidation and amino acid composition of the protein products led to their identification as NP-I-(9-93) (the 9-93 sequence) and [des-19,20]NP-I-(9-93) (the 9-93 sequence with the dipeptide 19-20 missing) for the more rapidly and more slowly formed species, respectively. NP-I-(9-93), unlike intact neurophysin I, is not retarded strongly by either Met-Tyr-Phe-amino-butylaminoagarose or neurophysin II-Sepharose. In contrast, both NP-I-(9-93) and [des-19,20]NP-I-(9-93) are equally as effective as intact NP-I in binding neurophysin I antibodies. The role of amino-terminal residues in promoting hormone binding, self-association, and antigenic recognition interactions is considered.

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Molecular, Thermodynamic, and Biological Aspects of Recognition and Function in Neurophysin‐Hormone Systems: A Model System for the Analysis of Protein–Peptide Interactions

Breslow

Burman

1990

Advances in Enzymology - And Related Areas of Molecular Biology

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Photoaffinity labeling of the hormone binding site of neurophysin.

Cited by 30 publications

References 37 publications

Dimerization of native and proteolytically modified neurophysins as monitored by proton magnetic resonance spectroscopy: proximity of tyrosine-49 to the subunit interface

Dimerization of native and proteolytically modified neurophysins as monitored by proton magnetic resonance spectroscopy: proximity of tyrosine-49 to the subunit interface

Effects of limited tryptic proteolysis of bovine neurophysins on molecular properties of hormone binding, self-association, and antigenicity

Molecular, Thermodynamic, and Biological Aspects of Recognition and Function in Neurophysin‐Hormone Systems: A Model System for the Analysis of Protein–Peptide Interactions

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