Abstract
The structure of the catalytically inactive mutant (C215S) of the human protein-tyrosine phosphatase IB (PTP1B) has been solved to high resolution in two complexes. In the first, crystals were grown in the presence of bis(-4nra-phosphophenyl)methane (BPPM), a synthetic high-affinity low molecular weight non-peptidic substrate (Km=l6 |iM), and the structure was refined to an R-factor of 18.2% at 1.9 A resolution. In the second, crystals were grown in a saturating concentration of phosphotyrosine (pTyr), and the structure was refined to an R-factor of 18.1 % at 1.85 A. Difference Fourier maps showed that BPPM binds FTP IB in two mutually exclusive modes, one in which it occupies the canonical pTyr-binding site (the active site), and another in which a phosphophenyl moiety interacts with a set of residues not previously observed to bind aryl phosphates. The identification of a second pTyr molecule at the same site in the PTPlB/C215S-pTyr complex confirms that these residues constitute a low-affinity non-catalytic aryl phosphatebinding site. Although the biological relevance of the second aryl phosphate binding site is unclear, its identification provides a new paradigm for the design of tight-binding, highly specific inhibitors for PTP1B.
| Original language | English (US) |
|---|---|
| Journal | FASEB Journal |
| Volume | 11 |
| Issue number | 9 |
| State | Published - 1997 |
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ASJC Scopus subject areas
- Agricultural and Biological Sciences (miscellaneous)
- Biochemistry, Genetics and Molecular Biology(all)
- Biochemistry
- Cell Biology
Cite this
Identincation of a novel aryl phosphate-binding site in ptp1b : implications for inhibitor design. / Zhan, Zhong Yin; Puius, Yoram A.; Zhao, Yu; Sullivan, Michael; Lawrence, David S.; Almo, Steven C.
In: FASEB Journal, Vol. 11, No. 9, 1997.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Identincation of a novel aryl phosphate-binding site in ptp1b
T2 - implications for inhibitor design
AU - Zhan, Zhong Yin
AU - Puius, Yoram A.
AU - Zhao, Yu
AU - Sullivan, Michael
AU - Lawrence, David S.
AU - Almo, Steven C.
PY - 1997
Y1 - 1997
N2 - The structure of the catalytically inactive mutant (C215S) of the human protein-tyrosine phosphatase IB (PTP1B) has been solved to high resolution in two complexes. In the first, crystals were grown in the presence of bis(-4nra-phosphophenyl)methane (BPPM), a synthetic high-affinity low molecular weight non-peptidic substrate (Km=l6 |iM), and the structure was refined to an R-factor of 18.2% at 1.9 A resolution. In the second, crystals were grown in a saturating concentration of phosphotyrosine (pTyr), and the structure was refined to an R-factor of 18.1 % at 1.85 A. Difference Fourier maps showed that BPPM binds FTP IB in two mutually exclusive modes, one in which it occupies the canonical pTyr-binding site (the active site), and another in which a phosphophenyl moiety interacts with a set of residues not previously observed to bind aryl phosphates. The identification of a second pTyr molecule at the same site in the PTPlB/C215S-pTyr complex confirms that these residues constitute a low-affinity non-catalytic aryl phosphatebinding site. Although the biological relevance of the second aryl phosphate binding site is unclear, its identification provides a new paradigm for the design of tight-binding, highly specific inhibitors for PTP1B.
AB - The structure of the catalytically inactive mutant (C215S) of the human protein-tyrosine phosphatase IB (PTP1B) has been solved to high resolution in two complexes. In the first, crystals were grown in the presence of bis(-4nra-phosphophenyl)methane (BPPM), a synthetic high-affinity low molecular weight non-peptidic substrate (Km=l6 |iM), and the structure was refined to an R-factor of 18.2% at 1.9 A resolution. In the second, crystals were grown in a saturating concentration of phosphotyrosine (pTyr), and the structure was refined to an R-factor of 18.1 % at 1.85 A. Difference Fourier maps showed that BPPM binds FTP IB in two mutually exclusive modes, one in which it occupies the canonical pTyr-binding site (the active site), and another in which a phosphophenyl moiety interacts with a set of residues not previously observed to bind aryl phosphates. The identification of a second pTyr molecule at the same site in the PTPlB/C215S-pTyr complex confirms that these residues constitute a low-affinity non-catalytic aryl phosphatebinding site. Although the biological relevance of the second aryl phosphate binding site is unclear, its identification provides a new paradigm for the design of tight-binding, highly specific inhibitors for PTP1B.
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UR - http://www.scopus.com/inward/citedby.url?scp=33750198648&partnerID=8YFLogxK
M3 - Article
AN - SCOPUS:33750198648
VL - 11
JO - FASEB Journal
JF - FASEB Journal
SN - 0892-6638
IS - 9
ER -