Preparation of Isotopically Labeled Recombinant Fragments of Fibronectin for Functional and Structural Study by Heteronuclear Nuclear Magnetic Resonance Spectroscopy - Methods in Molecular Biology: Extracellular Matrix Protocols - page 58

Fig. 1. 1 H- 13 C heteronuclear single quantum correlation (HSQC) spectrum of a
15 N/ 13 C-labeled sample of the fourth and fifth fibronectin type 1 module pair (8) ,
gathered using a 0.5 m M (5.5 mg/mL) sample. Using our fermentation procedure, 2 g
15 N-ammonium sulphate, 1 g U-[ 13 C]-glucose, and 20 g 13 C methanol were sufficient
to generate 60 mg pure 15 N / 13 C-labeled protein. The 1-dimensional projection
(A) of the 2-D matrix, (B) shows how overlapping peaks are resolved by extending
into the 13 C-dimension.
P. pastoris is not difficult, but we strongly advise anyone attempting it to
consult a fermentation scientist for general information about safe handling
and disposal; also there are variations in fermentation systems which may
necessitate small adaptations to our protocol.