Columns for protein purification

The cDNA for human kallistatin, human kallikrein-related peptidase 1 (KLK1) was purchased from GENECHEM. Fluorescent substrate H-D-Val-Leu-Arg-AMC for KLK1 was synthesized by BankPeptide. Low molecular weight (LMW) heparin with an average molecular weight of ≈5 kDa and high molecular weight (HMW) heparin with an average molecular weight of 17–19 kDa were purchased from Sigma. Heparin oligosaccharides with 4, 6, 8, 10, and 12 saccharide units (DP4, DP6, DP8, DP10, and DP12) were from Iduron (Manchester, UK). Columns for protein purification were purchased from GE Healthcare. All reagents and kits for crystallization were purchased from Hampton Research. Precast 12% SDS-PAGE gels were purchased from Genscript (Nanjing, China) and run at 150 volts for ≈ 45 min using MOPS buffer. Protein samples were treated with reducing SDS loading buffer at 95 °C for 5 min before electrophoresis.

If not stated otherwise, chemicals, antibiotics, and buffers were obtained from Sigma-Aldrich (St. Louis, MO) or Carl Roth GmbH (Karlsruhe, Germany); columns for protein purification were from GE Healthcare Life Sciences (Chicago, IL). The [9,10-³H] Triolein was obtained from PerkinElmer Life Sciences (Waltham, MA). Triolein, phosphatidylcholine, phosphatidylinositol, 1(rac)-oleoylglycerol, oleoyl-CoA, and free glycerol detection reagents were purchased from Sigma-Aldrich.