Sandbox323: Difference between revisions
From Proteopedia
Jump to navigationJump to search
No edit summary |
No edit summary |
||
| Line 5: | Line 5: | ||
4Q7Q is a homodimeric protein complex that originates from the bacterial species Chitinophaga Pinensis and has a mass of 58.5 kDa. It is a member of the SGNH Hydrolase Superfamily with structural and sequential similarities to esterases and lipases. Current evidence suggests it causes the hydrolysis of esters and/or acetyl groups on lipids/lipid-like molecules via a catalytic triad-like active site. | 4Q7Q is a homodimeric protein complex that originates from the bacterial species Chitinophaga Pinensis and has a mass of 58.5 kDa. It is a member of the SGNH Hydrolase Superfamily with structural and sequential similarities to esterases and lipases. Current evidence suggests it causes the hydrolysis of esters and/or acetyl groups on lipids/lipid-like molecules via a catalytic triad-like active site. | ||
<StructureSection load='4Q7Q' size=' | <StructureSection load='4Q7Q' size='200' side='right' caption='3D Representation of 4Q7Q's structure' scene=''> | ||
== Overview == | == Overview == | ||
| Line 80: | Line 80: | ||
=== Spectrophotometry === | === Spectrophotometry === | ||
[[Image:Ice_bath_basic_conditions_with_salt.jpg| | [[Image:Ice_bath_basic_conditions_with_salt.jpg|500px|left|]] | ||
50uL of purified protein from the first elution was added to 3mL of cold 1mg/mL PNPA. Solid sodium chloride was added to achieve a final concentration of 5mM NaCl. The absorbance peak was measured every 60 seconds for 30 minutes. Peak maximum was located at 427.8nm. The sample incubated in an ice bath between measurements. No discernible relationship between time and absorbance was noted. pH of the solution was 10.60. The instrument to measure pH was Vernier Lab Quest 3 with Tris-compatible flat pH sensor calibrated with two points. Theorizing the enzyme would increase activity at the physiological pH of common prokaryotes, we lowered the pH with 2M HCl. 2M NaOH was used to raise the pH due to the weak buffering capacity of the PNPA solution near pH 7. Final pH of the 1mg/mL PNPA solution was 7.30 | 50uL of purified protein from the first elution was added to 3mL of cold 1mg/mL PNPA. Solid sodium chloride was added to achieve a final concentration of 5mM NaCl. The absorbance peak was measured every 60 seconds for 30 minutes. Peak maximum was located at 427.8nm. The sample incubated in an ice bath between measurements. No discernible relationship between time and absorbance was noted. pH of the solution was 10.60. The instrument to measure pH was Vernier Lab Quest 3 with Tris-compatible flat pH sensor calibrated with two points. Theorizing the enzyme would increase activity at the physiological pH of common prokaryotes, we lowered the pH with 2M HCl. 2M NaOH was used to raise the pH due to the weak buffering capacity of the PNPA solution near pH 7. Final pH of the 1mg/mL PNPA solution was 7.30 | ||
