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The crystal structure of the triple mutant K53,56,120M of bovine pancreatic phospholipase A(2) has been redetermined using sulfur single-wavelength anomalous scattering. The synchrotron data were collected at lambda = 1.54 A and the crystal diffracted to 1.6 A resolution. The program SOLVE was used to locate the heavy atoms and to estimate the initial phases and the resulting map was then subjected to RESOLVE. The output of 455 non-H atoms, including 12 S atoms, one calcium ion and one chloride ion, were then subjected to ARP/wARP followed by REFMAC. With the improved phases, the automatic model building successfully built more than 85% of the 123 residues, excluding the N- and C-terminal residues. The final crystallographic R factor is 17.7% (R(free) = 21.7%). The refined model consists of 954 non-H protein atoms, 165 water O atoms, three 2-methyl-2,4-pentanediol (MPD) molecules, one calcium ion and one chloride ion. The present work is yet another example that shows the utility of single-wavelength anomalous scattering data for solving a protein structure.

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An 18-year old male Grant's Zebra Equus burchelli boehmi of Arignar Anna Zoological Park, Vandalur, developed lameness in the left forelimb due to overgrown hoof. The pastern region was swollen and the animal exhibited symptoms of pain during movement, and preferred to keep the affected limb off the ground. Parenteral anti-inflammatories and antibiotics were administered, but he showed little improvement. Wounds started to develop above the coronet, in the anterior and medial aspects and sero-sanguinous fluid started to ooze from the wounds. Because of the intense pain, the animal prefered to lie down and as a result, bed-sores developed over the elbow joints, shoulder and hock joints. The swelling extended further upward involving the lower third of the limb. Attempts for restraining manually proved futile and it was decided to chemically immobilize the zebra for detailed examination and treatment. Chemical immobilization:The zebra was immobilized with Etorphine hydrochloride (2.45mg) and Acepromazine maleate (10mg) using a blowpipe. Signs of anaesthesia were observed after 45 seconds as the animal tried to regain the standing posture with all the legs spread. The relaxed penis protruded and the head was held high after two minutes of the injection. The animal became sternally recumbent by the third minute and attained lateral recumbency after four minutes of injection. Observation and treatment:The affected limb was examined and the wounds were deep with thick pus, exposing the underlying second phalanx. The area was cleaned, the hoof was trimmed, the wounds were flushed with antiseptic solution, dry dressed and magnesium sulphate-glycerine paste applied over the swollen area and bandaged. A course of parenteral anti-inflammatory (Meloxicam @ 0.3mg/kg bodyweight i/m) and antibiotics (Amoxycillin + Cloxacillin @ 10mg/kg bodyweight i/ m) were administered.Revival from anaesthesia: Diprenorphine (3.26mg) was administered through intravenous route (45 minutes after induction). The animal attained sternal recumbency after two minutes, stood up after four minutes with relaxed penis and complete recovery was seen after 12 minutes.Nelson (1986) reported that hoof problems are commonly encountered in wild equids in all collections. The lameness in the present case was due to uneven hooves and old age, being the predisposing factor.

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