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Data
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Comments
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| Identiy of the enzyme |
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| Name of the reaction catalyst |
name, preferably the accepted name from the IUBMB Enzyme list |
| Fully balanced chemical reaction equation |
elements and charges balanced.
See convention in R.A. Alberty "Thermodynamics of Biochemical Reactions", doi:10.1002/0471332607 |
| EC number |
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| Oligomeric state |
number of different subunits |
| Sequence or sequence accession number |
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| Organism/species & strain |
NCBI Taxonomy ID |
| Additional information on the enzyme |
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| Isoenzyme |
naturally occuring variant or indication of selection of alternative |
| Tissue |
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| Organelle |
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| Localization |
within cell. Specify what localization is based on |
| Post-translational modification |
add only when determined |
| Preparation |
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| Description |
e.g., commercial source, procedure used or reference along with modifications |
| Artificial modification |
e.g., truncated, His-tagged, fusion protein, lacking native glycosylation |
| Enzyme or protein purity |
purity defined by which criteria. Specify whether protein or enzyme was purified. e.g., apparently homogeneous by PAGE, crude mitochondrial fraction, determined by MS |
| Metalloenzyme |
mutant, content, cofactors |
| Storage Conditions |
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| Storage temperature |
if frozen, freezing method, e.g., -20 °C flash |
| Atmosphere if not air |
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| pH |
e.g., 7.0 |
| At which temperature was the pH measured? |
e.g., 25 °C |
| Buffer & concentrations (including counter-ion) |
e.g., 200 mM potassium phosphate, 100 mM HEPES-KOH. If pH is adjusted by addition of acid or base not shown in the buffer name, make this clear - e.g., 50 mM sodium acetate adjusted with HCl. |
| Metal salt(s) & concentrations |
e.g., 10 mM KCl, 1.0 mM MgSO4 |
| Other components |
e.g., 1.0 mM EDTA, 1.0 mM dithiothreitol, 10% v/v glycerol, 20% w/w DMSO, 1 mg/ml PEG2000,
2 mg/ml BSA, peptidase inhibitors |
| Enzyme/protein concentration |
molar concentration if known, otherwise mass concentration;
e.g., µM or mg ml-1 |
Optional:
Statement about observed loss of activity under the above conditions |
e.g., less than 10% loss after 1 month |
| Statement about the thawing procedure |
e.g., on ice |
| Assay Conditions |
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| Identity and purity of all assay components |
identified unambiguously, ideally by reference to ID from database (such as PubChem, ChEBI), or using a textual identifier (such as InChI), and/or showing chemical structure (or SMILES).
Origin of compounds used with statement on purity. |
| Measured reaction |
as a stoichiometrically balanced equation
e.g., 2 mol substrate oxidized per mol O2 consumed, with all products identified |
| Assay temperature |
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| Assay pressure |
if it is not atmospheric; indicate if not aerobic |
| Atmosphere if not air |
|
| Assay pH |
How was it measured? |
| Buffer & concentrations |
e.g., 100 mM Tris-HCl, 200 mM potassium phosphate, including counter-ion.
If pH is adjusted by addition of acid or base not shown in buffer name, make this clear;
e.g., 50 mM sodium acetate adjusted with HCl. |
| Metal salt(s) & concentrations |
e.g., 10 mM KCl, 1.0 mM MgSO4 |
| Other assay components |
e.g., 1.0 mM EDTA, 1.0 mM dithiothreitol |
| Coupled assay components |
if relevant |
| Substrate & concentration ranges |
e.g., 1 - 100 mM glucose, 5 mM ATP |
| Enzyme/protein concentration |
molar concentration if known, otherwise mass concentration,
e.g., mg ml-1 or better: µM |
| Varied components |
e.g., inhibitor concentration |
| Total assay mixture ionic strength |
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| Activity |
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| Initial rates of the reaction measured |
determine how established, estimate ranges in substrate/product concentrations at the last data point.e.g., true initial tangent or average over specified time. |
| Proportionality between initial velocity and enzyme concentration |
if available |
| Enzyme activity |
Ideally kcat (i.e. Vmax/enzyme concentration), otherwise expressed as amount product formed per amount enzyme protein present per time unit.
Activity is sometimes reported as enzyme unit or international unit (1 IU = 1 µmol min-1). The katal (mol/s) may alternatively be used as a unit of activity (conversion factor 1 unit = 16.67 nkat).
Specify at which (range of) concentration(s) this was determined. |
Equilibrium measurements
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| Evidence that reaction reached equilibrium |
e.g., approached from both (or all if multiple substrates/products) directions |
| State which of all reactants were measured directly |
e.g., glucose phosphate and glucose measured directly, excess phosphate estimated by mass balance |
| Range of starting material and product concentrations in the experiment |
ideally a table of all initial and measured equilibrium concentrations |
| Complexing metal ions |
if the reaction involved species that might bind these (e.g., phosphate esters), essential to report estimated pMg and/or pCa |
| Methodology |
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| Assay method |
a literature reference may suffice for an established procedure but any modification should be detailed |
| Type of assasy |
e.g., continuous or discontinuous, direct or coupled |
| Reaction stopping procedure |
in the case of discontinuous assays |
| Direction of the assay |
with respect to the reaction equation provided,
e.g., NAD reduction by alcohol dehydrogenase;
alcohol + NAD+ → aldehyde or ketone + NADH + H+ |
| Reactant determined |
e.g., NADH formation, O2 utilization |
| Additional material desirable |
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| Free metal cation concentration |
e.g., of Mg2+ and Ca2+, specify how calculated |
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