1. Integral glycoprotein sequence, 195mer (Note amino acid sequence is broken into segments
containing 10 amino acids for ease in counting):
ZQMHNSDMGN (10) TKERYFLLVI (20) TLIFSFTILV (30) VAVYRASSRG[GFP] (40)
KHSDQRGDYF (50) FFIIILLLTS (60) VVVTAILYKE (70) S[BFP]NKHMYLSGG (80)
IAASTAAFGI (90) GLGGYKNED[BFP]L (100) KHWAIFFFFI (110) LVVVGALII (120)
GYEESQNSRE (130) NMRYIFHLPG (140) TTQNDDPPRN (150) TREKHNTSNT (160)
DESHMNQPPP (170) SSDHDKNPQT (180) YWYTSEKPLG (190) FKEEQ
Experimental comments:
A. Exposing the native membrane to an α-N-amino fluorescent labeling agent, demonstrated NO
retention of fluorescence within this specific membrane protein.
B. Exposing the native membrane to cyanogen bromide (CnBr), we were able to isolate a peptide
fragment that was carbohydrate positive.
C. Exposing the native membrane to tryptic digestion, we were NOT able to isolate a peptide
fragment that was carbohydrate positive.
D. The membrane protein DID NOT demonstrate FRET when exposed to 300 nm
Questions:
(i) Indicate the areas of sequence which would represent transmembrane spanning domains in a
native lipid bilayer
(ii) Draw a cartoon of the predicted transmembrane protein as it would be found in a native lipid
bilayer member – label all features (N- and C-termini), including carbohydrate positioning and
cleave sites used in defining protein orientation.
(iii) Draw a representative hydropathy plot for the above protein – Indicate the probable exit
points from