Difference between revisions of "Transformation of AA to 15-HPETE"

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(→‎Parameters)
(→‎15-LOX Parameters)
Line 25: Line 25:
 
|<math> mM </math>
 
|<math> mM </math>
 
|Human  
 
|Human  
−
|Epithelium
+
|Expression Vector: Epithelium
 +
Enzyme: 15-Lipoxygenase
 +
pH: 7.5
 +
Temperature: 25
 
|<ref name="Jacquot2008"> [http://www.ncbi.nlm.nih.gov/pubmed/18547056 Jacquot C. "Isotope sensitive branching and kinetic isotope effects in the reaction of deuterated arachidonic acids with human 12- and 15-lipoxygenases.'' Biochemistry. 2008 Jul 8;47(27):7295-303. doi: 10.1021/bi800308q. Epub 2008 Jun 12.]</ref>  
 
|<ref name="Jacquot2008"> [http://www.ncbi.nlm.nih.gov/pubmed/18547056 Jacquot C. "Isotope sensitive branching and kinetic isotope effects in the reaction of deuterated arachidonic acids with human 12- and 15-lipoxygenases.'' Biochemistry. 2008 Jul 8;47(27):7295-303. doi: 10.1021/bi800308q. Epub 2008 Jun 12.]</ref>  
 
|-
 
|-
Line 31: Line 34:
 
|<math> mM </math>
 
|<math> mM </math>
 
|Human  
 
|Human  
−
|Reticulocyte
+
|Expression Vector: Reticulocyte
 +
Enzyme: 15-Lipoxygenase
 +
pH:7.5
 +
Temperature: Unspecified
 
|<ref name="Jacquot2008b"> [http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2883171/ Jacquot C. "Synthesis of 11-Thialinoleic Acid and 14-Thialinoleic Acid, Inhibitors of Soybean and Human Lipoxygenases'' Org Biomol Chem. 2008 Nov 21; 6(22): 4242–4252.]</ref>  
 
|<ref name="Jacquot2008b"> [http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2883171/ Jacquot C. "Synthesis of 11-Thialinoleic Acid and 14-Thialinoleic Acid, Inhibitors of Soybean and Human Lipoxygenases'' Org Biomol Chem. 2008 Nov 21; 6(22): 4242–4252.]</ref>  
 
|-
 
|-
Line 37: Line 43:
 
|<math> mM </math>
 
|<math> mM </math>
 
|Human  
 
|Human  
−
|Keratinocyte
+
|Expression Vector: Keratinocyte
 +
Enzyme: 15-Lipoxygenase
 +
pH: 6.7 - 7.3
 +
Temperature: 37
 
|<ref name="Burrall1988"> [http://www.ncbi.nlm.nih.gov/pubmed/2459258 Burrall B. "Enzymatic properties of the 15-lipoxygenase of human cultured keratinocytes.'' J Invest Dermatol. 1988 Oct;91(4):294-7.]</ref>  
 
|<ref name="Burrall1988"> [http://www.ncbi.nlm.nih.gov/pubmed/2459258 Burrall B. "Enzymatic properties of the 15-lipoxygenase of human cultured keratinocytes.'' J Invest Dermatol. 1988 Oct;91(4):294-7.]</ref>  
 
|-
 
|-
Line 43: Line 52:
 
|<math> mM </math>
 
|<math> mM </math>
 
|Human
 
|Human
−
| Wildtype 15-Lipoxygenase
+
|Expression Vector: Baculovirus Insect Cell
 +
Enzyme: Wildtype 15-Lipoxygenase
 +
pH: 6.8
 +
Temperature: 37
 
|<ref name="Sloane1995"> [http://www.ncbi.nlm.nih.gov/pubmed/7479689 Sloane D. L. "Conversion of human 15-lipoxygenase to an efficient 12-lipoxygenase: the side-chain geometry of amino acids 417 and 418 determine positional specificity.'' Protein Eng. 1995 Mar;8(3):275-82..]</ref>  
 
|<ref name="Sloane1995"> [http://www.ncbi.nlm.nih.gov/pubmed/7479689 Sloane D. L. "Conversion of human 15-lipoxygenase to an efficient 12-lipoxygenase: the side-chain geometry of amino acids 417 and 418 determine positional specificity.'' Protein Eng. 1995 Mar;8(3):275-82..]</ref>  
 
|-
 
|-
Line 60: Line 72:
 
|<math> minute^{-1} </math>
 
|<math> minute^{-1} </math>
 
|Human  
 
|Human  
−
|Epithelium
+
|Expression Vector: Epithelium
 +
Enzyme: 15-Lipoxygenase
 +
pH: 7.5
 +
Temperature: 25
 
|<ref name="Jacquot2008"> [http://www.ncbi.nlm.nih.gov/pubmed/18547056 Jacquot C. "Isotope sensitive branching and kinetic isotope effects in the reaction of deuterated arachidonic acids with human 12- and 15-lipoxygenases.'' Biochemistry. 2008 Jul 8;47(27):7295-303. doi: 10.1021/bi800308q. Epub 2008 Jun 12.]</ref>  
 
|<ref name="Jacquot2008"> [http://www.ncbi.nlm.nih.gov/pubmed/18547056 Jacquot C. "Isotope sensitive branching and kinetic isotope effects in the reaction of deuterated arachidonic acids with human 12- and 15-lipoxygenases.'' Biochemistry. 2008 Jul 8;47(27):7295-303. doi: 10.1021/bi800308q. Epub 2008 Jun 12.]</ref>  
 
|-
 
|-
 
|37.2 ± 1.8
 
|37.2 ± 1.8
 
|rowspan="7"|per minute
 
|rowspan="7"|per minute
−
|rowspan="7"|Human prostate epithelial 15-lipoxygenase-2
+
|rowspan="7"||Expression Vector:Human prostate epithelial  
−
|pH 7
+
Enzyme:15-lipoxygenase-2
 +
|pH 7, Temperature: 22
 
|rowspan="7"|<ref name="Wecksler2009"> [http://www.ncbi.nlm.nih.gov/pubmed/18547056 Jacquot C. "Kinetic and Structural Investigations of the Allosteric Site in Human Epithelial 15-Lipoxygenase-2'' Biochemistry, 2009, 48 (36), pp 8721–8730]</ref>  
 
|rowspan="7"|<ref name="Wecksler2009"> [http://www.ncbi.nlm.nih.gov/pubmed/18547056 Jacquot C. "Kinetic and Structural Investigations of the Allosteric Site in Human Epithelial 15-Lipoxygenase-2'' Biochemistry, 2009, 48 (36), pp 8721–8730]</ref>  
 
|-
 
|-
 
|45 ± 1.2
 
|45 ± 1.2
−
|pH 7.5
+
|pH 7.5, Temperature: 22
 
|-
 
|-
 
|44.4 ± 2.4
 
|44.4 ± 2.4
−
|pH 8
+
|pH 8, Temperature: 22
 
|-
 
|-
 
|34.2 ± 0.6
 
|34.2 ± 0.6
−
|15°C
+
|15°C, pH = 7.5
 
|-
 
|-
 
|45 ± 1.2
 
|45 ± 1.2
−
|22°C
+
|22°C, pH = 7.5
 
|-
 
|-
 
|62.4 ± 4.2
 
|62.4 ± 4.2
−
|30°C
+
|30°C, pH = 7.5
 
|-
 
|-
 
|82.8 ± 4.8
 
|82.8 ± 4.8
−
|37°C
+
|37°C, pH = 7.5
 
|-
 
|-
 
|}
 
|}

Revision as of 16:23, 8 June 2016

Return to overview

Reaction

R17 AA - HPETE15.jpg

Chemical equation

AA \rightleftharpoons PGH2

Rate equation

15-LOX Parameters

Michaelis-Menten Constants
Value Units Species Notes Reference
3.70E-03 ± 3.00E-04 mM Human Expression Vector: Epithelium

Enzyme: 15-Lipoxygenase pH: 7.5 Temperature: 25

[1]
5.00E-03 mM Human Expression Vector: Reticulocyte

Enzyme: 15-Lipoxygenase pH:7.5 Temperature: Unspecified

[2]
1.06E-02 mM Human Expression Vector: Keratinocyte

Enzyme: 15-Lipoxygenase pH: 6.7 - 7.3 Temperature: 37

[3]
1.17E-02 ± 9.00E-04 mM Human Expression Vector: Baculovirus Insect Cell

Enzyme: Wildtype 15-Lipoxygenase pH: 6.8 Temperature: 37

[4]
Enzyme Turnover Numbers
Value Units Species Notes Reference
595.8 ± 16.8 minute^{-1} Human Expression Vector: Epithelium

Enzyme: 15-Lipoxygenase pH: 7.5 Temperature: 25

[1]
37.2 ± 1.8 per minute rowspan="7" Expression Vector:Human prostate epithelial

Enzyme:15-lipoxygenase-2

pH 7, Temperature: 22 [5]
45 ± 1.2 pH 7.5, Temperature: 22
44.4 ± 2.4 pH 8, Temperature: 22
34.2 ± 0.6 15°C, pH = 7.5
45 ± 1.2 22°C, pH = 7.5
62.4 ± 4.2 30°C, pH = 7.5
82.8 ± 4.8 37°C, pH = 7.5
Enzyme Concentrations
Value Units Species Notes Reference
1.60e-05 - 4.00e-05 mM Human Human reticulocyte 15-lipoxygenase-1, Enzymatic assays were performed in 25 mM Hepes buffer (pH 7.5,

25C), with and without product (12-HETE) addition, in the presence of 25 μM of substrate.

[5]
2.00e-05 mM Human Epithelium [1] _

References

  1. ↑ 1.0 1.1 1.2 Jacquot C. "Isotope sensitive branching and kinetic isotope effects in the reaction of deuterated arachidonic acids with human 12- and 15-lipoxygenases. Biochemistry. 2008 Jul 8;47(27):7295-303. doi: 10.1021/bi800308q. Epub 2008 Jun 12.
  2. ↑ Jacquot C. "Synthesis of 11-Thialinoleic Acid and 14-Thialinoleic Acid, Inhibitors of Soybean and Human Lipoxygenases Org Biomol Chem. 2008 Nov 21; 6(22): 4242–4252.
  3. ↑ Burrall B. "Enzymatic properties of the 15-lipoxygenase of human cultured keratinocytes. J Invest Dermatol. 1988 Oct;91(4):294-7.
  4. ↑ Sloane D. L. "Conversion of human 15-lipoxygenase to an efficient 12-lipoxygenase: the side-chain geometry of amino acids 417 and 418 determine positional specificity. Protein Eng. 1995 Mar;8(3):275-82..
  5. ↑ 5.0 5.1 Jacquot C. "Kinetic and Structural Investigations of the Allosteric Site in Human Epithelial 15-Lipoxygenase-2 Biochemistry, 2009, 48 (36), pp 8721–8730 Cite error: Invalid <ref> tag; name "Wecksler2009" defined multiple times with different content

Related Reactions

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