Difference between revisions of "Transformation of AA to 12-HPETE"

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(→‎12-LOX Parameters)
(→‎12-LOX Parameters)
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|<math> mM </math>
 
|<math> mM </math>
 
|Human  
 
|Human  
−
|Platelet
+
|Expression Vector: Platelet
 +
Enzyme: 12-Lipoxygenase
 +
pH: 7.4
 +
Temperature: 37°C.
 
|<ref name="Lagarde1984"> [http://www.ncbi.nlm.nih.gov/pubmed/6433902 Lagarde M. "Subcellular localization and some properties of lipoxygenase activity in human blood platelets.'' Biochem J. 1984 Sep 1;222(2):495-500.]</ref>   
 
|<ref name="Lagarde1984"> [http://www.ncbi.nlm.nih.gov/pubmed/6433902 Lagarde M. "Subcellular localization and some properties of lipoxygenase activity in human blood platelets.'' Biochem J. 1984 Sep 1;222(2):495-500.]</ref>   
 
|-
 
|-
Line 31: Line 34:
 
|<math> mM </math>
 
|<math> mM </math>
 
|Human  
 
|Human  
−
|Platelet
+
|Expression Vector: Platelet
 +
Enzyme: 12-Lipoxygenase
 +
pH: 7
 +
Temperature: 24
 
|<ref name="Hada1991"> [http://www.ncbi.nlm.nih.gov/pubmed/1851637 Hada T. "Catalytic properties of human platelet 12-lipoxygenase as compared with the enzymes of other origins.'' Biochim Biophys Acta. 1991 Apr 24;1083(1):89-93.]</ref>   
 
|<ref name="Hada1991"> [http://www.ncbi.nlm.nih.gov/pubmed/1851637 Hada T. "Catalytic properties of human platelet 12-lipoxygenase as compared with the enzymes of other origins.'' Biochim Biophys Acta. 1991 Apr 24;1083(1):89-93.]</ref>   
 
|-
 
|-
 
|1.00E-02
 
|1.00E-02
 
|<math> mM </math>
 
|<math> mM </math>
−
|Human
+
|Human Platlet
−
|Recombinant Histidine-Tagged Human Platelet 12-Lipoxygenase
+
|Expression Vector: Baculovirus
 +
Enzyme: 12-Lipoxygenase
 +
pH: 8
 +
Temperature: 37
 
|<ref name="Chen1993"> [http://www.ncbi.nlm.nih.gov/pubmed/8319693 Chen X. S. "Purification and characterization of recombinant histidine-tagged human platelet 12-lipoxygenase expressed in a baculovirus/insect cell system.'' Eur J Biochem. 1993 Jun 15;214(3):845-52.]</ref>   
 
|<ref name="Chen1993"> [http://www.ncbi.nlm.nih.gov/pubmed/8319693 Chen X. S. "Purification and characterization of recombinant histidine-tagged human platelet 12-lipoxygenase expressed in a baculovirus/insect cell system.'' Eur J Biochem. 1993 Jun 15;214(3):845-52.]</ref>   
 
|-
 
|-
Line 43: Line 52:
 
|<math> mM </math>
 
|<math> mM </math>
 
|Human  
 
|Human  
−
|Platelet 12-Lipoxygenase
+
|Expression Vector:  Platelets
 +
Enzyme:12-Lipoxygenase
 +
pH: 7.4
 +
Temperature: 37
 
|<ref name="Romano1993"> [http://www.ncbi.nlm.nih.gov/pubmed/8250832 Romano M. "Lipoxin synthase activity of human platelet 12-lipoxygenase.'' Biochem J. 1993 Nov 15;296 ( Pt 1):127-33.]</ref>   
 
|<ref name="Romano1993"> [http://www.ncbi.nlm.nih.gov/pubmed/8250832 Romano M. "Lipoxin synthase activity of human platelet 12-lipoxygenase.'' Biochem J. 1993 Nov 15;296 ( Pt 1):127-33.]</ref>   
 
|-
 
|-
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|per minute
 
|per minute
 
|Human
 
|Human
−
|Human reticulocyte 15-lipoxygenase-1, Enzymatic assays were performed in 25 mM Hepes buffer (pH 7.5,
+
|Expression Vector: Human reticulocyte
−
25C), with and without product (12-HETE) addition, in the presence of 25 μM of substrate.
+
Enzyme: 15-lipoxygenase-1
 +
pH: 7.5
 +
Temperature: 25
 
|<ref name="Wecksler2009"> [www.ncbi.nlm.nih.gov/pubmed/19469483 Wecksler A. "Mechanistic Investigations of Human Reticulocyte 15- and Platelet 12-Lipoxygenases with Arachidonic Acid'' Biochemistry, 2009, 48 (26), pp 6259–6267]</ref>
 
|<ref name="Wecksler2009"> [www.ncbi.nlm.nih.gov/pubmed/19469483 Wecksler A. "Mechanistic Investigations of Human Reticulocyte 15- and Platelet 12-Lipoxygenases with Arachidonic Acid'' Biochemistry, 2009, 48 (26), pp 6259–6267]</ref>
 
|-
 
|-
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|per minute
 
|per minute
 
|Wild Boar
 
|Wild Boar
−
|Wild Type Leukocyte-Type 12-Lipoxygenase Expressed in E. coli.
+
|Expression Vector: E. coli.
 +
Enzyme: 12-Lipoxygenase
 +
pH: 7.4
 +
Temperature: 37
 
|<ref name="Richards1997"> [http://pubs.acs.org/doi/abs/10.1021/bi963051a Richards K. "Leukocyte 12-Lipoxygenase:  Expression, Purification, and Investigation of the Role of Methionine Residues in Turnover-Dependent Inactivation and 5,8,11,14-Eicosatetraynoic Acid Inhibition'' Biochemistry, 1997, 36 (22), pp 6692–6699]</ref>
 
|<ref name="Richards1997"> [http://pubs.acs.org/doi/abs/10.1021/bi963051a Richards K. "Leukocyte 12-Lipoxygenase:  Expression, Purification, and Investigation of the Role of Methionine Residues in Turnover-Dependent Inactivation and 5,8,11,14-Eicosatetraynoic Acid Inhibition'' Biochemistry, 1997, 36 (22), pp 6692–6699]</ref>
 
|-
 
|-
Line 84: Line 101:
 
|mM
 
|mM
 
|Human
 
|Human
−
|Human reticulocyte 15-lipoxygenase-1, Enzymatic assays were performed in 25 mM Hepes buffer (pH 7.5,
+
|Expression Vector: Human reticulocyte
−
25C), with and without product (12-HETE) addition, in the presence of 25 μM of substrate.
+
Enzyme: 15-lipoxygenase-1
 +
pH: 7.5
 +
Temperature: 25
 
|<ref name="Wecksler2009"> [www.ncbi.nlm.nih.gov/pubmed/19469483 Wecksler A. "Mechanistic Investigations of Human Reticulocyte 15- and Platelet 12-Lipoxygenases with Arachidonic Acid'' Biochemistry, 2009, 48 (26), pp 6259–6267]</ref>
 
|<ref name="Wecksler2009"> [www.ncbi.nlm.nih.gov/pubmed/19469483 Wecksler A. "Mechanistic Investigations of Human Reticulocyte 15- and Platelet 12-Lipoxygenases with Arachidonic Acid'' Biochemistry, 2009, 48 (26), pp 6259–6267]</ref>
 
|-
 
|-

Revision as of 15:32, 8 June 2016

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Reaction

R179AA - HPETE12.jpg

Chemical equation

AA \rightleftharpoons PGH2

Rate equation

12-LOX Parameters

Michaelis-Menten Constants
Value Units Species Notes Reference
7.20E-03 mM Human Expression Vector: Platelet

Enzyme: 12-Lipoxygenase pH: 7.4 Temperature: 37°C.

[1]
8.00E-02 mM Human Expression Vector: Platelet

Enzyme: 12-Lipoxygenase pH: 7 Temperature: 24

[2]
1.00E-02 mM Human Platlet Expression Vector: Baculovirus

Enzyme: 12-Lipoxygenase pH: 8 Temperature: 37

[3]
7.90E-03 ± 8.00E-04 mM Human Expression Vector: Platelets

Enzyme:12-Lipoxygenase pH: 7.4 Temperature: 37

[4]
Enzyme Turnover Numbers
Value Units Species Notes Reference
336 ± 12 per minute Human Expression Vector: Human reticulocyte

Enzyme: 15-lipoxygenase-1 pH: 7.5 Temperature: 25

[5]
504 per minute Wild Boar Expression Vector: E. coli.

Enzyme: 12-Lipoxygenase pH: 7.4 Temperature: 37

[6]
Enzyme Concentrations
Value Units Species Notes Reference
5.00e-06 - 6.00e-05 mM Human Expression Vector: Human reticulocyte

Enzyme: 15-lipoxygenase-1 pH: 7.5 Temperature: 25

[5]

References

  1. ↑ Lagarde M. "Subcellular localization and some properties of lipoxygenase activity in human blood platelets. Biochem J. 1984 Sep 1;222(2):495-500.
  2. ↑ Hada T. "Catalytic properties of human platelet 12-lipoxygenase as compared with the enzymes of other origins. Biochim Biophys Acta. 1991 Apr 24;1083(1):89-93.
  3. ↑ Chen X. S. "Purification and characterization of recombinant histidine-tagged human platelet 12-lipoxygenase expressed in a baculovirus/insect cell system. Eur J Biochem. 1993 Jun 15;214(3):845-52.
  4. ↑ Romano M. "Lipoxin synthase activity of human platelet 12-lipoxygenase. Biochem J. 1993 Nov 15;296 ( Pt 1):127-33.
  5. ↑ 5.0 5.1 [www.ncbi.nlm.nih.gov/pubmed/19469483 Wecksler A. "Mechanistic Investigations of Human Reticulocyte 15- and Platelet 12-Lipoxygenases with Arachidonic Acid Biochemistry, 2009, 48 (26), pp 6259–6267]
  6. ↑ Richards K. "Leukocyte 12-Lipoxygenase:  Expression, Purification, and Investigation of the Role of Methionine Residues in Turnover-Dependent Inactivation and 5,8,11,14-Eicosatetraynoic Acid Inhibition Biochemistry, 1997, 36 (22), pp 6692–6699

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